Carnoy’s solution is a fixative composed of 60% ethanol, 30% chloroform and 10% glacial acetic acid, 1 gram of ferric chloride.^[1][2]

Carnoy’s solution is also the name of a different fixation composed of ethanol and glacial acetic acid (3:1).^[3][4][5]

Hm..and here are some articles that come up on a search for Carnoy’s solution at Science Direct I haven’t read them all but this may or may not be interesting:

Immunostaining Procedure

1. Fix the tissue samples immediately after surgical removal in 10% buffered formalin or in Carnoy’s solution (for chymase detection because formalin fixation destroys chymase immunoreactivity) for 12–24 hr at room temperature; then embed them in paraffin and cut into 4 μm thick sections. (this is step one out of twenty or so. I have an idea of what formalin is but no idea about chymase immunoreactivity so…rabbit holes.)

Handbook of Immunohistochemistry and in situ Hybridization of Human Carcinomas, Volume 3 Irene Esposito, … Helmut Friess, in Handbook of Immunohistochemistry and in Situ Hybridization of Human Carcinomas, 2005

• Chymase at Wikipedia (page last updated March 27, 2022)
  • Chymases (EC 3.4.21.39, mast cell protease 1, skeletal muscle protease, skin chymotryptic proteinase, mast cell serine proteinase, skeletal muscle protease) are a family of serine proteases found primarily in mast cells, though also present in basophil granulocytes (e.g. alpha chymase mcpt8). Recently, Derakhshan et al. reported that a specific mast cell population expressed trascripts for Mcpt8.^{[Derakhshan, Tahereh; Samuchiwal, Sachin K.; Hallen, Nils; Bankova, Lora G.; Boyce, Joshua A.; Barrett, Nora A.; Austen, K. Frank; Dwyer, Daniel F. (2021-01-04). “Lineage-specific regulation of inducible and constitutive mast cells in allergic airway inflammation”. Journal of Experimental Medicine. 218 (1): e20200321. doi:10.1084/jem.20200321. ISSN 0022-1007. PMC 7953627. PMID 32946563]} 
  • They show broad peptidolytic activity and are involved in a variety of functions. For example, chymases are released by mucosal mast cells upon challenge with parasites and parasite antigens promoting an inflammatory response, and chymase mcp1 and mcp2 are used for marker for mast cell degranulation in parasite infection such as: 
    • Nematode,^{[McDermott JR, Bartram RE, Knight PA, Miller HR, Garrod DR, Grencis RK (June 2003). “Mast cells disrupt epithelial barrier function during enteric nematode infection”. Proceedings of the National Academy of Sciences of the United States of America. 100 (13): 7761–6. Bibcode:2003PNAS..100.7761M. doi:10.1073/pnas.1231488100. PMC 164661. PMID 12796512]} 
    • Trichuris muris^{[Betts CJ, Else KJ (January 1999). “Mast cells, eosinophils and antibody-mediated cellular cytotoxicity are not critical in resistance to Trichuris muris”. Parasite Immunology. 21 (1): 45–52. doi:10.1046/j.1365-3024.1999.00200.x. PMID 10081771. S2CID 31343469][Blackwell NM, Else KJ (June 2001). “B cells and antibodies are required for resistance to the parasitic gastrointestinal nematode Trichuris muris”. Infection and Immunity. 69 (6): 3860–8. doi:10.1128/IAI.69.6.3860-3868.2001. PMC 98409. PMID 11349052.]} 
  • Chymases are also known to convert angiotensin I to angiotensin II and thus play a role in hypertension and atherosclerosis.^{[Caughey GH (June 2007). “Mast cell tryptases and chymases in inflammation and host defense”. Immunological Reviews. 217: 141–54. doi:10.1111/j.1600-065X.2007.00509.x. PMC 2275918. PMID 17498057.]}
  • Because of its role in inflammation it has been investigated as a target in the treatment of asthma.^{[de Garavilla L, Greco MN, Sukumar N, Chen ZW, Pineda AO, Mathews FS, et al. (May 2005). “A novel, potent dual inhibitor of the leukocyte proteases cathepsin G and chymase: molecular mechanisms and anti-inflammatory activity in vivo”. The Journal of Biological Chemistry. 280 (18): 18001–7. doi:10.1074/jbc.M501302200. PMID 15741158]}
  • Categories: 
    • Genes on human chromosome 14
    • EC 3.4.21
    • Hydrolase stubs

Endopeptidases: serine proteases/serine endopeptidases (EC 3.4.21)

Enzymes

• Google suggests Chymosin and Chymase are synonyms so I’m going to have to make a new page in attempt to make better sense of this gobbledygook..

Carnoy’s Solution Uses

Some of the uses of Carnoy’s solution are:

• Enhancing lymph node detection during dissection of cadavers.^[6]
• Immunohistochemical fixation and detection of NMDA receptors within the murine hippocampus.^[7]
• Applied directly following enucleation for the treatment of odontogenic keratocysts.^[8][9][10]
• Direct application following enucleation (Cuba) for certain kinds of unicystic ameloblastomas.^[11] This appears to decrease the likelihood of recurrence over enucleation alone.^[12] Protein coagulation is thought to limit uptake of these toxic materials by surrounding tissues, however it is this fact that limits its usefulness as a treatment agent in general.^[13]
• As a fixative for pap smear samples.^[14]
• As a fixative agent for both nuclear and mitochondrial DNA in various tissues.^[15]
• As a fixative agent to preserve mucus, useful for tissue preparation before staining with periodic acid-Schiff base.^[16]

Karyotyping

The routine chromosome analysis (Karyotyping) refers to analysis of metaphase chromosomes which have been banded using trypsin followed by Giemsa, Leishmanns, or a mixture of the two. This creates unique banding patterns on the chromosomes. The molecular mechanism and reason for these patterns are unknown, although it likely related to replication timing and chromatin packing.

Several chromosome-banding techniques are used in cytogenetics laboratories. Quinacrine banding (Q-banding) was the first staining method used to produce specific banding patterns. This method requires a fluorescence microscope and is no longer as widely used as Giemsa banding (G-banding). Reverse banding, or R-banding, requires heat treatment and reverses the usual black-and-white pattern that is seen in G-bands and Q-bands. This method is particularly helpful for staining the distal ends of chromosomes. Other staining techniques include C-banding and nucleolar organizing region stains (NOR stains). These latter methods specifically stain certain portions of the chromosome. C-banding stains the constitutive heterochromatin, which usually lies near the centromere, and NOR staining highlights the satellites and stalks of acrocentric chromosomes.

High-resolution banding involves the staining of chromosomes during prophase or early metaphase (prometaphase), before they reach maximal condensation. Because prophase and prometaphase chromosomes are more extended than metaphase chromosomes, the number of bands observable for all chromosomes (bands per haploid set, bph; “band level”) increases from about 300 to 450 to as many as 800. This allows the detection of less obvious abnormalities usually not seen with conventional banding.^{[ Geiersbach, Katherine B.; Gardiner, Anna E.; Wilson, Andrew; Shetty, Shashirekha; Bruyère, Hélène; Zabawski, James; Saxe, Debra F.; Gaulin, Rebecca; Williamson, Cynthia; Van Dyke, Daniel L. (February 2014). “Subjectivity in chromosome band–level estimation: a multicenter study”. Genetics in Medicine. 16 (2): 170–175. doi:10.1038/gim.2013.95.]}

Slide preparation

Cells from bone marrow, blood, amniotic fluid, cord blood, tumor, and tissues (including skin, umbilical cord, chorionic villi, liver, and many other organs) can be cultured using standard cell culture techniques in order to increase their number. A mitotic inhibitor (colchicine, colcemid) is then added to the culture. This stops cell division at mitosis which allows an increased yield of mitotic cells for analysis. The cells are then centrifuged and media and mitotic inhibitor are removed, and replaced with a hypotonic solution. This causes the white blood cells or fibroblasts to swell so that the chromosomes will spread when added to a slide as well as lyses the red blood cells. After the cells have been allowed to sit in hypotonic solution, Carnoy’s fixative (3:1 methanol to glacial acetic acid) is added. This kills the cells and hardens the nuclei of the remaining white blood cells. The cells are generally fixed repeatedly to remove any debris or remaining red blood cells. The cell suspension is then dropped onto specimen slides. After aging the slides in an oven or waiting a few days they are ready for banding and analysis.

Analysis

Analysis of banded chromosomes is done at a microscope by a clinical laboratory specialist in cytogenetics (CLSp(CG)). Generally 20 cells are analyzed which is enough to rule out mosaicism to an acceptable level. The results are summarized and given to a board-certified cytogeneticist for review, and to write an interpretation taking into account the patient’s previous history and other clinical findings. The results are then given out reported in an International System for Human Cytogenetic Nomenclature 2009 (ISCN2009)..

References

1. ^ “MSDS :: Carnoy’s Solution (Fixative)”. Archived from the original on 2009-03-27. Retrieved 13 Jan 2009.
2. ^ Carnoy J. B. (1887). “Appendice Les Globule Polaires de L’Ascaris Clavata”. La Cellule RECUEIL DE CYTOLOGIE ET d’HISTOLOGIE GÉNÉRALE. 3: 276.
3. ^ Tjio JH, Whang J (1962). “Chromosome Preparatons of Bone Marrow Cells without PriorIn VitroCulture orIn VivoColchicine Administration”. Stain Technology. 37: 17–20. doi:10.3109/10520296209114563. PMID 13921436.
4. ^ Mazia D, Brewer PA, Alfert MA (1953). “The Cytochemical Staining and Measurement of Protein with Mercuric Bromphenol Blue”. The Biological Bulletin. 104 (1): 57–67. doi:10.2307/1538691. JSTOR 1538691.
5. ^ Utevsky S, Kovalenko N, Doroshenko K, Petrauskienė L, Klymenko V (2009). “Chromosome numbers for three species of medicinal leeches (Hirudo spp.)”. Systematic Parasitology. 74 (2): 95–102. doi:10.1007/s11230-009-9198-2. PMID 19731093. S2CID 7947757.
6. ^ Luz DA, Ribeiro U, Chassot C, Collet E, Silva FS, Cecconello I, Corbett CE (December 2008). “Carnoy’s solution enhances lymph node detection: an anatomical dissection study in cadavers”. Histopathology. 53 (6): 740–2. doi:10.1111/j.1365-2559.2008.03148.x. PMID 19076686. S2CID 19362456.
7. ^ Yoneyama M, Kitayama T, Taniura H, Yoneda Y (August 2003). “Immersion fixation with Carnoy solution for conventional immunohistochemical detection of particular N-methyl-D-aspartate receptor subunits in murine hippocampus”. J. Neurosci. Res. 73 (3): 416–26. doi:10.1002/jnr.10622. PMID 12868075. S2CID 40884078.
8. ^ Madras J, Lapointe H (March 2008). “Keratocystic odontogenic tumour: reclassification of the odontogenic keratocyst from cyst to tumour”. J Can Dent Assoc. 74 (2): 165–165h. PMID 18353202.
9. ^ “Odontogenic Keratocyst: The Northwestern USA Experience”. Archived from the original on 2009-01-06. Retrieved 14 Jan 2009.
10. ^ “Use of Carnoy’s Solution in management of odontogenic keratocysts”. Archived from the original on 2009-05-17. Retrieved 14 Jan 2009.
11. ^ Lee PK, Samman N, Ng IO (April 2004). “Unicystic ameloblastoma–use of Carnoy’s solution after enucleation”. Int J Oral Maxillofac Surg. 33 (3): 263–7. doi:10.1006/ijom.2003.0496. PMID 15290793.
12. ^ Lau SL, Samman N (August 2006). “Recurrence related to treatment modalities of unicystic ameloblastoma: a systematic review”. Int J Oral Maxillofac Surg. 35 (8): 681–90. doi:10.1016/j.ijom.2006.02.016. PMID 16782308.
13. ^ Marx, Robert E.; Stern, Diane (2003). Oral and maxillofacial pathology: a rationale for diagnosis and treatment. Chicago: Quintessence. p. 684. ISBN 0-86715-390-3.
14. ^ Shamsi M, Abdali K, Montazer NR, Kumar PV, Tabatabaee HR (2008). “Comparison of Carnoy’s solution and 96% ethyl alcohol fixation in bloody Pap smears”. Acta Cytol. 52 (2): 187–90. doi:10.1159/000325477. PMID 18499991. S2CID 3370408.
15. ^ Miething F, Hering S, Hanschke B, Dressler J (March 2006). “Effect of fixation to the degradation of nuclear and mitochondrial DNA in different tissues”. J. Histochem. Cytochem. 54 (3): 371–4. doi:10.1369/jhc.5B6726.2005. PMID 16260588.
16. ^ “Stains File: Carnoy’s Fluid”. Archived from the original on 15 December 2010. Retrieved 25 Oct 2009.

Categories: 

• Histology
• Pathology
• Laboratory techniques
• Medical terminology

From Wikipedia where this page was last modified 19 February 2022,