[Histonet] immuno diluent vs. buffer question
I have a question for those doing immunos by hand. I typically dilute my primary, secondary and tertiary antibodies with manufacturer supplied diluent. Recently, I was running a test to determine the source of a mysterious precipitate that had begun to appear on my slides (It has since then disappeared just as mysteriously). In one of the conditions, I substituted the diluent with TBST for all dilutions. I found that this particular slide had noticeably less background. When inquiring as to what I should be diluting these reagents with, I received conflicting answers from tech support. I heard that the secondary and tertiary antibodies should be diluted with what they are actually in, i.e. .05% TBS or 1M PBS. I also was told that I could dilute everything with diluent. Can anyone tell me which is correct? Or why using the buffer yielded less background than the diluent? Aside from sodium azide and BSA I don't know what's in the proprietary reagent. This communication is intended solely for the use of the addressee and may contain information that is legally privileged, confidential or exempt from disclosure. If you are not the intended recipient, please note that any dissemination, distribution, or copying of this communication is strictly prohibited. Anyone who receives this message in error should notify the sender immediately and delete it from his or her computer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] barcode labels vs handwritten
Hi Folks, I am running into resistance with a few of our pathologists regarding the use of barcoded labels. They want us to handwrite our slides and then apply our permanent labels over top of that. We are using the Stainershield labels that are solvent-resistant. We eliminated the step of relabeling our slides with the permanent labels at the sorting area, and began having the cutter label their slides with the permanent labels at the microtome area as part of a Lean workflow initiative. However, a few of our Pathologists like having the handwriting underneath the label so they can see who cut the slide if it is of poor quality. Also, if they think it is mislabeled they say that gives them a clue as to where the slide really came from. I guess I feel like, if the slide is mislabeled it doesn't matter where it came from, it still needs to be recut from the correct block. I'd appreciate your comments and suggestions about this issue. Thanks, Angie Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] ?bacterial/fungal contamination on slides
Fungi can grow in a number of different aqueous stain solutions. One time a tech brought me a slide she had stained with GMS for fungi, together with the positive control slide she had run. I could see fungi on the test slide but there was no sign of silver staining. The control slide showed positively stained fungi as expected, but also the same unstained fungi seen on the test slide. It turned out a fungus was growing in the aqueous solution of Fast Green we used as a counterstain for the GMS. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Humidity levels in the lab
Julie, I can't offer you any references, but here's something to consider. A cool mist humidifier in your lab. We are in Bend, OR., on the dry side of the Cascades. Running the humidifier reduces static and makes sectioning a bit easier. No hard science with this, just a little more atmospheric moisture. By the way, the humidifier I bought is a big blue and white penguin. The mist streams out the beak. Just looking at it makes you smile. I bought it at Target for ~ $40.00 - it works for us. Have a good weekend. Tom J. Thomas Jasper HT (ASCP) BAS Histology Supervisor Central Oregon Regional Pathology Services Bend, Oregon 97701 541/693-2677 [EMAIL PROTECTED] -Original Message- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Randolph-Habecker, Julie Sent: Thursday, September 18, 2008 3:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Humidity levels in the lab Folks, We just moved to a new lab space in a new building. We are currently running at 45% or lower humidity. I am working with our facilities on raising the level but I am looking for some references for what level we should aim for in regard to paraffin cutting. Does anyone have some ideas or places to look for information? Thanks!! Julie Julie Randolph-Habecker, Ph.D. Staff Scientist - Director Experimental Histopathology Shared Resource Fred Hutchinson Cancer Research Center 1100 Fairview Ave, N. DE-360 (Please note new location) Seattle WA 98109-1024 206-667-6119 [EMAIL PROTECTED] ___ 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
