Supravital staining is a method of staining used in microscopy to examine living cells that have been removed from an organism. It differs from intravital staining, which is done by injecting or otherwise introducing the stain into the body. Thus a supravital stain may have a greater toxicity, as only a few cells need to survive it a short while. The term "vital stain" is used by some authors to refer specifically to an intravital stain, and by others interchangeably with a supravital stain, the core concept being that the cell being examined is still alive. As the cells are alive and unfixed, outside the body, supravital stains are temporary in nature.[1][2]
Supravital stain of a smear of human blood from a patient with hemolytic anemia. The reticulocytes are the cells with the dark blue dots and curved linear structures (reticulum) in the cytoplasm.
The most common supravital stain is performed on reticulocytes using new methylene blue or brilliant cresyl blue, which makes it possible to see the reticulofilamentous pattern of ribosomes characteristically precipitated in these live immature red blood cells by the supravital stains. By counting the number of such cells the rate of red blood cell formation can be determined, providing an insight into bone marrow activity and anemia.[3] This is in contrast to vital staining, when the dye employed is one that is excluded from the living cells so that only dead cells are stained positively. (Vital stains include dyes like trypan blue and propidium iodide, which are either too bulky or too charged to cross the cell membrane, or which are actively rapidly pumped out by live cells.)
Supravital staining can be combined with cell surface antibody staining (immunofluorescence) for applications such as FACS analysis.[4] Immunofluorescence can also be done within the interior of live cells by reversible cell permeabilization using the detergent Triton X-100. Adjusted carefully to the appropriate concentration for the number of cells, the pretreatment can permit access of molecules between 1 and 150 kilodaltons to the interior of the cell.[5] Although antibodies may be used in a similar way in this context, the term "supravital stain" is typically reserved for smaller chemicals which possess suitable properties intrinsically.
^N. Chandler Foot (1954). "Vital staining". Annals of the New York Academy of Sciences. 59 (2): 259–267. doi:10.1111/j.1749-6632.1954.tb45936.x. PMID 13229212.
^Tina Carlsen; Richard Langlois; Erin Bissell. "Immunoassay/Vital Stain System (IVSS) Proof-of-Concept Final Report" (PDF). Lawrence Livermore National Laboratory.
^Anne L van de Ven, Karen Adler-Storthz, and Rebecca Richards-Kortum (2009). "Delivery of Optical Contrast Agents using Triton-X100, Part 1: Reversible permeabilization of live cells for intracellular labeling". J Biomed Opt. 14 (2): 021012. doi:10.1117/1.3090448. PMC2748244. PMID 19405725.{{cite journal}}: CS1 maint: multiple names: authors list (link)
^2011 Hematology, Clinical Microscopy, and Body Fluids Glossary by CAP
April 10, 2024
supravital, staining, method, staining, used, microscopy, examine, living, cells, that, have, been, removed, from, organism, differs, from, intravital, staining, which, done, injecting, otherwise, introducing, stain, into, body, thus, supravital, stain, have, . Supravital staining is a method of staining used in microscopy to examine living cells that have been removed from an organism It differs from intravital staining which is done by injecting or otherwise introducing the stain into the body Thus a supravital stain may have a greater toxicity as only a few cells need to survive it a short while The term vital stain is used by some authors to refer specifically to an intravital stain and by others interchangeably with a supravital stain the core concept being that the cell being examined is still alive As the cells are alive and unfixed outside the body supravital stains are temporary in nature 1 2 Supravital stain of a smear of human blood from a patient with hemolytic anemia The reticulocytes are the cells with the dark blue dots and curved linear structures reticulum in the cytoplasm The most common supravital stain is performed on reticulocytes using new methylene blue or brilliant cresyl blue which makes it possible to see the reticulofilamentous pattern of ribosomes characteristically precipitated in these live immature red blood cells by the supravital stains By counting the number of such cells the rate of red blood cell formation can be determined providing an insight into bone marrow activity and anemia 3 This is in contrast to vital staining when the dye employed is one that is excluded from the living cells so that only dead cells are stained positively Vital stains include dyes like trypan blue and propidium iodide which are either too bulky or too charged to cross the cell membrane or which are actively rapidly pumped out by live cells Supravital staining can be combined with cell surface antibody staining immunofluorescence for applications such as FACS analysis 4 Immunofluorescence can also be done within the interior of live cells by reversible cell permeabilization using the detergent Triton X 100 Adjusted carefully to the appropriate concentration for the number of cells the pretreatment can permit access of molecules between 1 and 150 kilodaltons to the interior of the cell 5 Although antibodies may be used in a similar way in this context the term supravital stain is typically reserved for smaller chemicals which possess suitable properties intrinsically Examples of common supravital dyes editSupravital dyes for RBCs 6 New methylene blue Brilliant cresyl blue Crystal violet Methyl violet Nile blueHoechst stainReferences edit English Medical Dictionary N Chandler Foot 1954 Vital staining Annals of the New York Academy of Sciences 59 2 259 267 doi 10 1111 j 1749 6632 1954 tb45936 x PMID 13229212 John P Greer Maxwell Myer Wintrobe 2008 Wintrobe s clinical hematology ISBN 9780781765077 Tina Carlsen Richard Langlois Erin Bissell Immunoassay Vital Stain System IVSS Proof of Concept Final Report PDF Lawrence Livermore National Laboratory Anne L van de Ven Karen Adler Storthz and Rebecca Richards Kortum 2009 Delivery of Optical Contrast Agents using Triton X100 Part 1 Reversible permeabilization of live cells for intracellular labeling J Biomed Opt 14 2 021012 doi 10 1117 1 3090448 PMC 2748244 PMID 19405725 a href Template Cite journal html title Template Cite journal cite journal a CS1 maint multiple names authors list link 2011 Hematology Clinical Microscopy and Body Fluids Glossary by CAP Retrieved from https en wikipedia org w index php title Supravital staining amp oldid 994685588, wikipedia, wiki, book, books, library,
