Hi Charles A method my EM scientist used many years ago was quite simplistic and he thought it worked well. You simply centrifuge the specimen, add a set volume of deionised water, mix to lose the red cells, then add an equal volume of double strength normal saline to create a suspension in normal saline. The centrifuge where the heavier uncleared cells go to the bottom of the tube. I have to say the I have never tried this personally but if the morphology was retained for EM it must be fine for LM. Having said that I have great respect for Tony Henwood and his method looks much more scientific.
Regards Tony Sent from my iPhone > On 17 Jan 2020, at 8:24 am, Tony Henwood (SCHN) via Histonet > <histonet@lists.utsouthwestern.edu> wrote: > > Hi Jennifer, > > I have had excellent success with lysing the red blood cells (using Isotonic > Ammonium Chloride) prior to cell block preparation with > thromboplastin-plasma. > The lysing solution contains EDTA so you will need to add a few drops of 1% > calcium chloride. Method as follows: > > Lysis solution > Ammonium Chloride 4.5g > Potassium carbonate 0.5g > EDTA 0.0186g > Distilled water 500mls > > Method: > 1. Centrifuge bloody fluid. > 2. Remove supernatant and add equal volume of lysis solution. > 3. Resuspend and incubate for 5 minutes at 4oC. > 4. Centrifuge, if blood still remains, then repeat from step 2. > 5. Rinse in Hanks or RPMI, centrifuge. > 6. Mix pellet in a few drops of plasma. > 7. Add thromboplastin and a few drops of 1% Calcium Chloride, mix gently > and allow clot to form. > 8. Add 10% buffered formalin and fix and process as usual. > > Reference: > Kuenen-Boumiester etal (1996) Acta Cytolog 40:475-479 > > If you donot use the plasma clot method for cell block preparation, then use > your preferred method after step 5. > The lysis solution can also be purchased commercially from several companies > (eg Biolegend). It is commonly used for sample preparation for flow > cytometry. Check the SDS to make sure it does not contain formaldehyde. > > > -----Original Message----- > From: Mac Donald, Jennifer via Histonet > [mailto:histonet@lists.utsouthwestern.edu] > Sent: Friday, 17 January 2020 4:53 AM > To: Charles Riley <cri...@dpspa.com>; Histo List > <histonet@lists.utsouthwestern.edu> > Subject: Re: [Histonet] Cell block preparations > > Acetic acid would work. > > Get Outlook for iOS<https://aka.ms/o0ukef> ________________________________ > From: Charles Riley via Histonet <histonet@lists.utsouthwestern.edu> > Sent: Thursday, January 16, 2020 8:55:21 AM > To: Histo List <histonet@lists.utsouthwestern.edu> > Subject: [Histonet] Cell block preparations > > EXTERNAL SENDER- Exercise caution with requests, links, and attachments. > > What is the best way to remove excess blood from FNA sample collections > before spinning them down into cell blocks? > > -- > > Charles Riley BS HT, HTL(ASCP)CM > > Histopathology Coordinator/ Mohs > _______________________________________________ > 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 > > This message is intended for the addressee named and may contain confidential > information. If you are not the intended recipient, please delete it and > notify the sender. > > Views expressed in this message are those of the individual sender, and are > not necessarily the views of NSW Health or any of its entities. > > > _______________________________________________ > 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
