What is the procedure for staining microscopic laboratory slides for
1) biopsies
2) pap smears
3) blood smears
4) are there others?
Answer should provide very specific detail that includes A) the type
of solution, B) the concentration of the solution (e.g. isopropyl
alcohol 70%) and the amount of time left in a solution. Please
include information about rinsing using tap or distilled water if
applicable.
The answer should provide the above information for the first three
categories and any additional categories that might be applicable.
Sources for the information would be helpful.Hello lakeerieboater,
There are a plethora of tinctorial staining techniques available,
and I have collected links for you, to numerous commonly used staining
techniques that outline the solutions, concentrations, and the type of
water used. In a hospital and/or research lab, one can assume
distilled water for rinsing is intended. One would not want *any*
possible contaminants or an inconsistent pH to interfere with staining
procedures. Some of the following techniques do state ?tap? water
however, these being less sensitive stains. Because of
protection, I am unable to copy the procedures. Instead, I am
providing you with sites that describe in detail the staining process.
After completing most of this answer, I came across the following
comprehensive site that lists practically every known biological stain
currently in use, along with solutions, concentrations, procedures and
what type of water is required: tap water vs. DI H20.
http://members.pgonline.com/~bryand/StainsFile/stain/stainindex.htm
Please be aware that in most labs today, a majority of these staining
techniques are now automated, especially blood film staining. Some
lesser run and esoteric tests, such as a smear for eosinophils, or
stool for white blood cells are still stained manually, in some
laboratories, as are many bacterial slides. Some staining techniques
simply don?t adapt well to automation, and are still done manually, as
in smears for fetal blood cell presence in maternal blood.
The two most commonly used stains in a clinical lab would be Wright
Stain (for blood differentials) and Gram stain (for bacteria). A
research lab?s most commonly used stains would depend on the area of
research,
1)Biopsies:
======================================
The stain of choice would depend on what one is looking for in a biopsy:
Pancreas Tissue stain: GOMORI'S ALDEHYDE FUCHSIN and several others:
http://home.primus.com.au/royellis/ST%5CPANCREAS.htm
Mast Cells in tissue
Acidified toluidine blue and chloroacetate techniques:
http://home.primus.com.au/royellis/ST%5CMASTCELL.htm
Mast cells, acidic polysaccharides, sulfomucins, sialomucins and
uronic acid mucins in tissue.
http://www.biostains.org/new_page_16.htm
Amyloid (Connected with Down's syndrome, Alzheimer's disease and the
tra nsmissible spongiform encephalopathies. .Congo Red and Mayer?s
techniques. http://home.primus.com.au/royellis/amyloid.htm
Mitochondria (Altmann?s Stain and Van Gieson's Stain)
http://www.biostains.org/featured_dye.htm
Tissue for acid fast organisms, such as tuberculosis
http://www.imebinc.com/stainkits/procedures/afb.html
Tissue for bacteria:
Brown & Brenn Tissue Gram Stain
Carazzi?s hematoxylin
http://www.nottingham.ac.uk/pathology/protocols/carazz.html
Connective Tissue Stains
http://www.biostains.org/aniline_blue.htm
Sudan Black (For fats)
http://members.pgonline.com/~bryand/StainsFile/stain/lipid/chiffelleputtpropgly.htm
Numerous tissue stains can be found on this page:
http://home.primus.com.au/royellis/haem.html
2) Pap smears
There are numerous different Pap stain formulas.
This is a bought, prepared stain:
http://www.gbi-inc.com/Cyto.htm
Another Pap stain (Note, this one does use tap water!):
http://www.agora.crosemont.qc.ca/urinesediments/doceng/doc_034.htm
Yet another Pap stain:
http://members.pgonline.com/~bryand/StainsFile/stain/nuclei/llewmb3.htm
This page shows a picture of an automated Pap smear stainer (bottom of page)
http://www.pamf.org/pathology/qualitybefore.html
3) Blood smears
Routine blood films utilize Wright stain
http://www.tpub.com/content/medical/14295/css/14295_286.htm
Page 4 of this document describes a method for blood film staining,
and malarial smear staining. Page 5 contains the procedure for
reticulocyte staining.
http://static.cjp.com/gems/blood/LH.6.1.Houwen.pdf
Simple blood stain for malarial parasites (Tap water used)
http://www.malariajournal.com/content/1/1/2
Romanowsy Stain (Often used to highlight cell nuclei, platelets and parasites)
http://home.primus.com.au/royellis/ST%5CROMTECH.htm
Reticulocytes in blood smears (Methlylene blue)
http://w3.whosea.org/haem/c12.htm
4) Others
Bone Marrow (And other tissue, for fungus). PAS(Periodic acid Schiff )
and other methods
http://www5d.biglobe.ne.jp/~hasumi/method/pas_e.html
Urine:
I could not find the formula for Sternheimer-Malbin stain
(crystal-violet and safranin), a stain used for urine sediment. This
stain is usually not used by experienced technologists, but is
available in most labs, and comes already prepared, in a dropper
bottle.
http://www.vh.org/adult/provider/pathology/CLIA/UrineAnalysis/3.1BPreparing.html
Hank?s Stain (Urine casts, cells)
http://www.agora.crosemont.qc.ca/urinesediments/doceng/doc_031.htm
Bile:
Fouchet?s stain for presence of bile
http://www.hoslink.com/histo/8.HTM#8.3_bile
Nasal secretions, urine, tissue:
Hansel?s Stain (for eosinophils, and can distinguish allergies from
colds on a nasal secretion specimen)
Bacteria:
Gram Stain:
http://www.ncl.ac.uk/dental/oralbiol/oralenv/tutorials/gramstain.htm
http://www.wfcc.info/tis/info9.html
http://www.glue.umd.edu/~nsw/ench485/lab9b.htm#List
No specifics as to staining solutions, but a good overview of the Gram Stain:
http://www.spjc.edu/hec/vettech/vtde/ATE2639LGS/gramstain.htm
Ziehl-Nielsen stain (Mycobacteria, aspergillosis, more)
http://www.studentbmj.com/back_issues/0404/education/142.html
Geimsa
http://www.nottingham.ac.uk/pathology/protocols/mgg.html
Fungus:
http://www.nottingham.ac.uk/pathology/protocols/grocott.html
http://service.merck.de/microbiology/tedisdata/nodes/5073.html
Stool Parasites:
Gomori Tri-chrome stain for parasites (Requires DI H20):
http://www.med-chem.com/Procedures/Wheatleys%20TRICHROME%20STAIN.pdf
This site details a modified acid-fast stain, a trichrome stain and
several others for parasites:
http://www.dpd.cdc.gov/dpdx/HTML/DiagnosticProcedures.asp?body=Frames/DiagnosticProcedures/body_dp_stoolstaining.htm
Myelin (Covering of nerves):
http://www.nottingham.ac.uk/pathology/protocols/weil.html
Alizarin Red S (bone) (Scroll down to near middle of page)
http://home.primus.com.au/royellis/BONE.htm
METHYL METHACRYLATE, and other bone stains can also be found on the above page.
Lugol?s Iodine (plant biology and tissue-vaginal cancer)
http://regentsprep.org/Regents/biology/units/laboratory/indicators.cfm
http://www.path.org/files/RH_vili.pdf
http://www.syndel.com/msds/lugol_msds.html
Methylene Violet for presence of yeast, in beermaking!
http://www.asbcnet.org/Journal/abstracts/search/1999/0204-03a.htm
Many stains today are bought already prepared:
http://www.bd.com/clinical/products/stains/stainmyco.asp
More stain information:
http://www.lab.anhb.uwa.edu.au/mb140/MoreAbout/stains.htm
Commonly used stains:
http://www.sigmaaldrich.com/Brands/Sigma/Hematology_Histology/Certified_Stains_and_Dyes.html
http://www.uq.edu.au/vdu/HDULeukaemiaCytochem.htm
I hope this adequately answered your question. Again, please request
an Answer Clarification, if any part of my answer requires further
explanation, and I will be glad to assist you further, if possible.
Regards,
crabcakes
biochemical stains
biological stain methods
hematological stain techniques
tissue staining proceduresThank you for the stars and nice tip, lakeerieboater. Both are appreciated.
Sincerely,
crabcakes#If you have any other info about this subject , Please add it free.# |
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8 January 2009 |
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