[Histonet] Immunofluorescense staining
I just want to ask about precipitation reaction of secondary antibody especially on goat antibody conjugate Alexa dye 594 and 488 Invitrogen. I have noticed that the method I used using 1% BSA and 3% BSA blocking , then overnight with primary has reacted to my secondary goat anti Rabbit and mouse alexa dyre 594 and 488 with too much precipitation. I tried to remove it with PBS with 0.05 tween wash overnight and the precipitate are still there. Any help would be appreciated with my technique. Thank you so much. Reuel Cornelia Cellular Pathology Laboratory Supervisor Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel. Number: 214-559-7766 FAX Number: 214-347-4806 Scottish Rite for Children is a world-renowned leader in the treatment of orthopedic conditions such as scoliosis, clubfoot, hand differences, hip disorders, sports injuries and fractures, as well as certain related arthritic and neurological disorders and learning disorders, such as dyslexia. Patients receive treatment regardless of the family's ability to pay. For more information about services available at our Dallas or Frisco campuses, visit scottishriteforchildren.org. This email transmission and/or its attachments may contain confidential health information, intended only for the use of the individual or entity named above. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Multiplex IHC
Anyone doing multiplex IHC. I have not heard of it until today. Any reference or recommendations with regards to instrumentation and protocol would be highly appreciated. Thank you. Reuel Cornelia Cellular Pathology Laboratory Supervisor Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel. Number: 214-559-7766 FAX Number: 214-347-4806 Scottish Rite for Children is a world-renowned leader in the treatment of orthopedic conditions such as scoliosis, clubfoot, hand differences, hip disorders, sports injuries and fractures, as well as certain related arthritic and neurological disorders and learning disorders, such as dyslexia. Patients receive treatment regardless of the family's ability to pay. For more information about services available at our Dallas or Frisco campuses, visit scottishriteforchildren.org. This email transmission and/or its attachments may contain confidential health information, intended only for the use of the individual or entity named above. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Oil Red O
I was wondering if you could help me know why our Oil Red O have some black snowflakes on our fat tissue hours after staining or after 24 hours more snowlflakes precipitation occurs. The staining was reference was from Lillie RD, Ashburn. Please note that our staining works well between two to three hours. Is there a reason fro this precipitation? Reuel TSRH Dallas, TX -- Texas Scottish Rite Hospital for Children is one of the nation's leading pediatric centers for the treatment of orthopedic conditions, certain related neurological disorders and learning disorders, such as dyslexia. This email transmission and/or its attachments may contain confidential health information, intended only for the use of the individual or entity named above. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Depalstify
Hello histonetters, Regarding my first email. The thickness of my section is 5 micron. Reuel I have a problem with my MMA plastic section on my slide. I could not remove the plastic MMA even if I leave the section slides in Xylene for several days with heat incubation at 60 degrees. This is the first time that happened to my section for several years of doing the same thing over. I was thinking that it was my Xylene lot number so I tried to change it but still does not remove the plastic, and then Itried different solvent Acetone, and Ethylene glycol Monoethyl ether but still it does not work. Can I ask for help if anyone knows what was going on and what would be the best way to remove this plastic from my section? Just to give you my embedding solution was MMA -94% , dibutly phthalate- 5% and perkadox 16- 0.5%. This have been my solution for years and I do not have any problem with the removal of plastic section until now. I was thinking that my MMA (M55909) different lot number from Sigma Aldrich may have cause this because even I tried to use the MMA to dissolve my plastic it does not work. Thank you for and any opinions or protocols are greatly appreciated. Reuel Cornelia TSRH 214-559-7766 -- Texas Scottish Rite Hospital for Children is one of the nation's leading pediatric centers for the treatment of orthopedic conditions, certain related neurological disorders and learning disorders, such as dyslexia. This email transmission and/or its attachments may contain confidential health information, intended only for the use of the individual or entity named above. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Deplastifying MMA
Hello histonetters, I have a problem with my MMA plastic section on my slide. I could not remove the plastic MMA even if I leave the section slides in Xylene for several days with heat incubation at 60 degrees. This is the first time that happened to my section for several years of doing the same thing over. I was thinking that it was my Xylene lot number so I tried to change it but still does not remove the plastic, and then Itried different solvent Acetone, and Ethylene glycol Monoethyl ether but still it does not work. Can I ask for help if anyone knows what was going on and what would be the best way to remove this plastic from my section? Just to give you my embedding solution was MMA -94% , dibutly phthalate- 5% and perkadox 16- 0.5%. This have been my solution for years and I do not have any problem with the removal of plastic section until now. I was thinking that my MMA (M55909) different lot number from Sigma Aldrich may have cause this because even I tried to use the MMA to dissolve my plastic it does not work. Thank you for and any opinions or protocols are greatly appreciated. Reuel Cornelia TSRH 214-559-7766 -- Texas Scottish Rite Hospital for Children is one of the nation's leading pediatric centers for the treatment of orthopedic conditions, certain related neurological disorders and learning disorders, such as dyslexia. This email transmission and/or its attachments may contain confidential health information, intended only for the use of the individual or entity named above. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Non Specific Esterase
Hello Histonetters, Does anyone have a protocol for NSE on paraffin sections? Reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Reference to microtome micrometer thickness
Hello Histonetters, how will we verify the thickness of a paraffin section in a slide. Do we have a reference regarding on how to measure the thickness. I based the thickness of my section thru the mcirometer on the rotary microtome but one of our reveiwers does not believed that we are cutting them into 3 micron thickness or 4 micron thickness. Please if we have any reference, please share it to me. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Squamous cells staining on H&E and IHC
Wear gloves and do not scratch your head or nose with gloves on. >>> "Margiotta-Watz, Michele" 10/29/2014 8:57 AM >>> We have that problem mostly during the winter months when our hands get dry. Use hand lotion, that usually helps a bit. Thanks, Michele Margiotta-Watz Histology Supervisor BMHMC 101 Hospital Rd. Patchogue, NY 11772 631-654-7192 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amber McKenzie Sent: Tuesday, October 28, 2014 3:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Squamous cells staining on H&E and IHC Does anyone else have problems with what looks like squamous cells staining on your H&E's and IHC's? I'm trying to figure out how to eliminate that problem in our lab...wear gloves while cutting? Change out water bath several times during shifts? Any suggestions? Thanks! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] CD68
Does anybody out there knows a CD68 antibody that works with pig(Porcine) tissue. We have tried 3 CD68 antibodies already from Santa Cruz, Dako and Novus that they say works with pig tissue but it did not. We tried using heat antigen retrieval and enzyme digestion for paraffin, it did not work. We also use pig frozen tissue and still do not work. Any suggestion. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Perfusion of mouse bone femur
Does anybody have a Protocol for perfusion of mouse bone femur. It looks like my protocol does not work. 1.fix in 4% PFA in PBS for 24 hrs in refrigerator 2. wash in PBS 3x 3. transfer in 14 %EDTA for 5 days. 4. Wash in water for maximum of 2 hrs in running water 6. perfusion with 15% sucrose overnight 7.perfusion with 30% overnight. 8. Transfer in OCT. 8. Cut in 10 um and transfer section on APES slide Results: When cutting section, the tissue does not adhere to the slides properly. Can you please help us or any comment to correct this protocol. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Thermo Scientific STP 420ES Tissue Processor
Hello histonetter, Do you have any feedback on the efficiency of the Thermo Scientific STP 420ES Tissue Processor. We are planning to purchase a tissue processor that will meet our goal in reducing the time of bone tissue processing that we use both for plastic(MMA) and paraffin research and clinical tissue. We will be having our demo instrument on tomorrow. Any feedback, or opinion regarding this processor will be of great help. Thank you. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Tissue Culture No telephone regulation
Hello everyone,, I wanted to know what is the rules and regulation requirement for not having a telephone inside a tissue/Cell culture room. I have been told that we could not placed a telephone inside a culture room. But their are equipment like microscope attached to a PC, incubators or simply a call for emergency that needs access to a phone for communications. Does a telephone can create a big mess of contamination to a tissue culture room? I tried to find a policy to verify that a telephone cannot be installed inside a tissue culture room but NIH and other facility says that you just have to clean the surfaces of the phone and computer. I am not a user of the culture room but it has been so difficult to make communication when something goes wrong with the equipment inside that room since I take care of making the communication and how does the safety of the staff inside be assured if in case there is a possible fire or simply fainting inside. Please help me find this policy and I will stand to what is regulated. Thnak you very much. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Substitute Xylene for Bone tissue
Does the Xylene substitute work very well with large bone tissue (5 to 7 cm long and between 3-5 mm thickness) for MMA and routine processing. What will be out pitfalls? We have a staff in our lab who is so "sensitive" to the smell of Xylene as she wanted to get pregnant sometime and the little smell that we have in our lab makes her emotional about it. Our lab has a very good ventilation but you could not prevent the smell when you open a retort after processing or even wiping small residue and placing it in our biohazard bin or simply cover slipping using xylene base mountant. I was planning to try the substitute to help make our environment a better place for everyone. Your helpful advise is always appreciated. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Thanks!
This was a good discussion and a some little rough comment but we learn from it. I have read all the feedback on xylene and its substitute and it gives us a lesson to become more better histotechs. Just this morning one of our administrator passed by near our processor while my co worker has just opened the processor retort for cleaning and smelled xylene in the air. He stopped and asked us why he was not using a mask while doing that. I replied that was only a minimal amount of xylene in the retort that was left after the cleaning cycle and he said well I just want to make sure of your safety. When we are so used of the smell sometimes we do not think of our Safety and this discussion taught us again to think first of our own Safety whether you are sensitive or not to xylene or any chemical. To shelly, I wish your employee the best and all success in the field of histology. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 >>> "D'Attilio, Shelley" 9/28/2011 11:16 AM >>> Hi all, I wanted to thank all of you who responded to my question about my xylene-sensitive employee and purchase of an air filter system. I got tons of terrific information and enjoyed the discussion about xylene substitutes. Those who responded gave me a lot to think about and raised some questions for me to have answered. It's wonderful to have access to all of you experts on the listserv! Thank you for your generosity. Regards, Shelley D'Attilio MT(ASCP) Manager, Chemistry, Cytology and Histology Dept. of Pathology and Laboratory Medicine Stormont-Vail HealthCare Topeka, Kansas NEED A DOCTOR? Stormont-Vail's Health Connections can help you find a doctor accepting new patients. Call (785) 354-5225. ** The information transmitted in this e-mail and in any replies and forwards are for the sole use of the above individual(s) or entities and may contain proprietary, privileged and/or highly confidential information. Any unauthorized dissemination, review, distribution or copying of these communications is strictly prohibited. If this e-mail has been transmitted to you in error, please notify and return the original message to the sender immediately at the above listed address. Thank you for your cooperation. ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Plastic embedding in dallas area
Does anybody in Dallas,TX area are working on plastic embedding that are charging them. Please let me know because I have a student from UTA who are working on fiber optic on nerve tissue that wants to do plastic embedding. Our facility does not work on outside samples unless we are in collaboration. Thank you for your help. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MMA
Can undecalcified bone MMA embedded tissue be use for Electron Microscopy.The tissue was fixed in formalin dehydrated in grades of alcohol, clear in xylene, infiltrated and embedded in MMA (MMA, dibutyl phthalate,perakdox). If not,can anyone have a procedure how to prepare tissue that are embedded in MMA for EM. Is this the same procedure done on a paraffin embedded tissue where you melt the paraffin with xylene then hydrate, wash in distilled water then transfer in osmium ,wash in water, dehydrate, PO,resin. Thank you for your help. Reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MMA for EM
Can MMA embedded tissue be use for Electron Microscopy. If not,can anyone have a procedure how to prepare tissue that are embedded in MMA for EM. Is this the same procedure done on a paraffin embedded tissue. Thank you for your help. Reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Joplin's article
To those who were not able to open to the site, I tried to copy and paste and hopefully you can read it. This is a very interesting story about Joplin's St John's Regional Medical Center and code Gray. If you still have problems. please visit this link. http://m.kansascity.com/kcstar/db_41535/contentdetail.htm?contentguid=0kA2g6z2&src=cat&full=true#display By ERIC ADLER and LAURA BAUER The Kansas City Star JOPLIN, Mo. | Looking back, they remember the quiet — like a last, deep breath before death. In the nursery of St. John’s Regional Medical Center, newborns napped in bassinets. Ventilators hummed in an intensive-care unit. In the emergency room, nurse Tracy Hernandez checked an older woman for a stroke, one of the few serious cases in the ER all day. In an operating room down a second-floor hallway, orthopedic surgeon James “Dusty” Smith opened an infected hip. One floor above, John Seay, a 60-year-old mechanic from Welch, Okla., visited with his 83-year-old mother. Frail from congestive heart failure, she doubted she’d be getting better and had picked out a pink dress for her burial. Then, in the west, the air began to spin. Condition Gray. The announcement over the hospital speakers warned of a potential tornado. Prepare. No one panicked. Such calls are routine in Joplin, a zinc- and lead-mining town carved from the rock and fields of Tornado Alley. Nurses pulled shades over windows to shield from flying debris. They rolled equipment from the halls, on the off chance patients would have to be moved there. Off chance, because this storm wasn’t expected to hit them. Visitors watched it on television with nonchalance. Radar showed funnel clouds tracking north. In a neighborhood across from St. John’s, Amanda and Bradley German sat in a friend’s home with their sons, Brody, 6, and James, 9, heedless of the weather alert. Small hail fell. The friend tossed hailstones playfully into the house. “We were joking about it,” Amanda German would say. “We hear the storm sirens all the time.” What no one anticipated was the dark monster developing to the west, two miles outside their windows. The sky turned the green of a violent bruise. Execute Condition Gray: Get patients to safety! • • • It’s been four weeks since an EF-5 tornado slammed to the ground in Joplin, its 200-mph winds scouring a three-quarter-mile-wide, six-mile-long band of devastation. When it was over, this city of 50,000 would reel, broken and bloodied. For the watching world, the image of the hollowed shell of St. John’s was ground zero. For 115 years, the hospital system created by the Catholic order of the Sisters of Mercy healed the community’s illnesses and injuries. In seconds, the nine-story symbol of the city’s strength and caring, built and expanded since 1968, would stand gravely injured itself. Yet in many ways, the story of what happened inside the walls of St. John’s might better stand as a microcosm of the horrors, heroism and humanity that played out across Joplin that night. Witness upon witness recounts a stream of “walking wounded,” individuals impaled by wood, glass or metal, limbs missing, flesh torn from their bodies, lurching toward the hospital. Many remain haunted by the carnage they saw. But that night at the hospital, they said, also will be remembered as one of the city’s proudest. There was the surgeon who operated by flashlight as the hospital crashed around him. The ER doc who plunged a chest tube through the ribs of a young man to keep him from dying. Nurses used their bodies to blanket vulnerable patients from wind-hurled debris. A floor tech plucked a flying man from the air. Employees wielded axes to free drugs from locked cabinets. And then there were the strangers. Hundreds rushed in convoys of pickup trucks, descending on the hospital to speed the wounded away. Hospital visitors left the sides of their dead and dying loved ones to carry fragile patients down blackened hallways, guided by the dim light of cell phones. “We did what we had to do,” said John Seay. • • • At 5:41 p.m., the tornado descends, a black, twisting wall on the western horizon. It splinters houses, strips trees, heaves cars. Hailstones crash through glass like sledgehammers. Rain pounds down in a stinging curtain. Minutes away, St. John’s waits — 183 patients in its 367 licensed beds; some 25 patients in the ER; 100 staff on duty; an unknown number of visitors in the patient rooms, halls and waiting areas. In the ER, Angie Abner, 40 — a paramedic who became a nurse only a year ago, and who has missed work the last two days because of food poisoning — has been doing triage. Now, minutes after the call Execute Condition Gray, she is struggling to get patients to safety, into hallways and away from windows. People, having grown too used to these warnings and then seeing storms peter out, refuse to move. “Folks, this is for your own safety,” Abner belts, emphatic. “You have to listen to me!” A man waves
[Histonet] Fwd: Article on the Tornado in the Joplin hospital
A must read article on code Gray. Very sad story but heroic. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histotech needed in dallas texas
Research - Histotechnologist This position is responsible for supporting research projects in histological techniques, immunohistochemistry, hard tissue histology and image analysis of pathological specimens with Molecular biology, confocal microscopy and laser capture microdissection background. B.S. in medical technology/histology or B.S. in biomedical sciences required. HT (ASCP) or eligible preferred, with a minimum of five years work experience. M-F, 8 a.m. - 4:30 p.m. For more information or to apply on-line, please visit us at www.tsrhc.org or contact Shonna Norman at tsr...@tsrh.org. Texas Scottish Rite Hospital for Children Welborn Street Dallas, Texas 75219 214-559-7590 Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cryojane Model 400
I just want to know if anybody out there in histoland have an Instrumedics cryojane Model 400 that is more than 10 years old and the spare parts like the UV bulb, heating pad are still supported by a company. Leica just told me this afternoon that they do not support any spare parts of an instrument more than 10 years old. Please give me your feedback. Thank you. Reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cryojane expertise
I need help with cryojane techniques on bone tissue.I have a hard time picking up the bone trabeculae area and bone marrow but the cartilage and other soft tissue are so easy to pick up. Any advice on the technique? Thank you. Reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Laser Capture Machine
To anybody using a laser capture machine, I want to ask your opinion on the following machine which one is better with regard to their instrumentation and cost of consumables because we are in the process of purchasing one and your honest opinion with regards to handling this machine is very much accountable. 1. Arcturus XT 2. Leica AS LMD 3. P.A.L.M LPC 4. MMI Cell Cut 5. Arcturus PixCell lle Thank you very much. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Test for cell viability in paraffin section?
Would you know a techniqe to confirm cell viability in paraffin embedded tissue? Propidium and methylene blue staining are used in cell culturing to detect cell viability. Is there similar technique for detecting cell viability in tissue? This question was asked by our research director, Can you please share your opinion on this and if there is chemical test to detect vaibility of cells in a paraffin tissue? Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MMP9 and MMP13 on pig tissue
Does anybody out there working on MMP9 and 13 on Pig tissue. Where do you purchase your antibody. Thank you. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] how to prevent foldings on femoral head cartilage tissue
Hello histonetters especailly hard tissue group I have a pig femoral head bone tissue embedded in paraffin and I have a hard time getting rid of the folding problem. I tried to remedy by lowering my temperature to 38 C and putting them in 5% alcohol before placing them in water bath I still have a lots of folding formation on some areas of the cartilage. Is there any other technique to remedy this problem. I appreciate your help. Thank you. reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Large fibrous Bone
I would like to thank everyone for their contribution on how to remedy our large fibrous bone tissue. Just for your knowledge, this case is a Congenital Pseudoathrosis(CPT) and we have process them both for paraffin and MMA. The paraffin was the choice for our study since we will be doing a lot of IHC studies on this. I was able to cut this hard fibrous tissue by placing them longer(1 hr) on ice and constanly surface decal (30minutes) even if a complete decal was achieve. Thank you histonetters for your unselfish knowledge and experience. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] large fibrous bone tissue
How do you process a fibrous bone tissue ( 7 mm thick). We have use Paraffin Type 9 from Richard allan Scientific to embed works well with our bone femur( 7 mm) when cutting but on fibrous bone it does not give us a good result in cutting the blocks. It is like cutting a uterus tissue but a little bit harder. Please give me your opinion on how to remedy this kind of tissue not mentioning double embedding method or plastic. Thank you. Reuel Cornelia ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Double staining of the same Primary
Can anybody share their protocol for Rat spinal cord double immunostain both flourescent and ABC-HRP of the same species (mouse) Primary antibody. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Pig tissue IHC detection
I just wanted to know if anybody who are working with Pig bone tissue fix in 10%NBF and decalcified in 14% EDTA, citrate buffer (dako) antigen retrieval pH7.0 and ph 9.0 by steaming for 20 minutes, what is their IHC detection kit for mouse monoclonal and rabbit polyclonal antibodies. I am using a Powervision kit from Immunovision, now Leica and I have a lot of background staining even with my negative stain. Please help me remedy the background stain.Thank you. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC detection for Pig tissue?
I just wanted to know if anybody who are working with Pig bone tissue fix in 10%NBF and decalcified in 14% EDTA, citrate buffer (dako) antigen retrieval pH7.0 and ph 9.0 by steaming for 20 minutes, what is their IHC detection kit for mouse monoclonal and rabbit polyclonal antibodies. I am using a Powervision kit and I have a lot of background staining even with my negative staining. Please help. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] CD 31 on Pig
Is anyone using CD 31 on Pigs. Where do you purchase your antibody. Thank you. Reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Bone tissue
We have a difficulty cutting metatarsal bone . It seems that our sections are so dried up. I was thinking that our dehydration have something to do with this which we have placed it in a wrong processing procedure for our large bone. The tissue is 4 mm thick and 1-2 cm in length and width and was dehydrated in 70% - 4 hrs, 80%-4 hrs,95% -4 hrs and 2 changes of 100% 3 hrs each, paraffin is 4 hrs each 2 changes. The tissue was decalcified in 14% EDTA. When we start cutting them it is so brittle and we could not even create a section. I have surfaced decal it and also place in a softener Mollifex some of it work but some does not work. Please help us remedy this tissue. Thank you. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Slide for bone tissue
Dear histonetters, I need to know an available slides for bone tissue especcally with cartillage. I have done a positively charged slides and did a subbed slides but it fails on our special staining and immunohistochem. Please help us. Does APES slides will help resolved this problem? Thank you. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Non specific Esterase protocol
Can somebody share a Non specific esterase Enzyme hsitochemistry protocol that works well with frozen muscle tissue. I have a protocol that I have used for ages but it does not give me a very strong reddish brown staining on my angular fibers. I will be happy to have it and compare. Thank you. Reuel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] bone tissue
Can somebody help me with my bone tissue to remain intact during antigen retrieval for IHC staining. I have used subbed slides but It did not help me. Is there more better adhesive. Does Haupts Gelatin works better? I know this is a hundred year old question and hopefully this problem have been resolved since my undated knowledge could remember. I would like to thank everybody for responding to my last e-mail on gross photography. It gives me more idea what to purchase. Thanks again histonetters and I really appreciate your help. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Gross photography/macrophotography
Hello histonetters, I am looking for a good digital camera for gross photography. Any recommendations that works with your lab will be beneficial. Thank you. Happy memorial day!!! Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: RE: [Histonet] IHC on paraformaldehyde-fixed
I have been curious about this discussion. we used 4% paraformaldehyde for smaller biopsies only because it has a faster penetration to tissue than 10% formalin. In all my IHC that I have done. I observe that doing an IHC with 4% paraformaldehyde does not necessarily need antigen retrieval in comparison to 10% formalin either it will be human or animal tissue but this depends on how long was it fix, our 4% paraformaldehyde we fix smaller biopsies like nerve,muscle, skin for 6 to 12 hrs. and for formalin it is 12 to 48 hours or more. Maybe you can comment on the effect on this to tissue if you say you will use 4% paraformaldehyde for storage and transportation. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 >>> "Tony Henwood" <[EMAIL PROTECTED]> 12/04/08 9:29 PM >>> tf wrote: "I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~." I have not heard this before. Do you have a reference for this? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: tf [mailto:[EMAIL PROTECTED] Sent: Friday, 5 December 2008 2:11 PM To: Tony Henwood; [EMAIL PROTECTED]; Jan Shivers; histonet Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed the basic principles are the same for most cross-linking fixatives and induce similar bonds the difference you observed between may due to any other variability, or the co-fixative you used. I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~. 2008-12-05 tf 发件人： Tony Henwood 发送间： 2008-12-05 06:00:03 收件人： [EMAIL PROTECTED]; Jan Shivers; histonet 抄送： 主： RE: [Histonet] IHC on paraformaldehyde-fixed Interesting point. Since 10% buffered formalin (made from the concentrated 38% formaldehyde) contain about 1% methanol, has it been shown that this has a deleterious effect on ANY antigens or are we expecting this worse case senario as being the norm? I am not aware of any antigens (or antigen-antibody combination) that has been badly effected by 10% formalin that is NOT effected by 10% formaldehyde. Are you aware of any?? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] Sent: Friday, 5 December 2008 1:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -Original Message- From: Tony Henwood <[EMAIL PROTECTED]> Date: Thu, 04 Dec 2008 09:35:09 To: Jan Shivers<[EMAIL PROTECTED]>; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is
