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UND 363 Alc. blu hya
UND 363 Alcian Blue w/ Hualuronidase
Question | Answer |
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Purpose of Alc. blu. Hyaluron | to differintiate epithelial and connective tissue mucins |
Principle of Alc. blu. Hyaluron | testicular enzyme hyaluronidase will digest out the mucal polysaccharides found in connective tissue, therefore epithelial tissue will still stain while connective tissue will show diminished or NO stain |
Fixative for Alc. blu. Hyaluron | 10% NBF |
Technique requirements for Alc. blu. Hyaluron | 2 identical sections of umbilical cord @ 4-5 microns (on 2 slides) one labeled with (digestion) and one W/O (for connective tissue mucin) |
what is an alternative technique with Alc. blu. Hyaluron (for epithelial mucins) | 2 identical sections/ 2 slides (W/ and W/O) of sm. bowel, appendix, colon (to demonstrate epithelial mucins) |
Microscope for Alc. blu. Hyaluron | light |
name the 4 major reagents in Alc. blu. Hyaluron | 1. Buffer (both slides get the buffer but one will get the enzyme) 2. hyaluronidase digestion solution 3. alcian blue (stain mucin) 4. nuclear fast red (for counterstain) |
explain in detail the buffering step | on slide is put in only buffer, the other is put in digestion solution w/ buffer. They are give the same time and temp. Before staining they are put in 3% acetic acid to prevent PH change of the soln' |
What are the standard results of Alc. blu. Hyaluron | W/O digestion - acid mucopolysaccharidase and sialomucins will be deep blue W/ digestion mucosubstances containing hyaluronic acid and chondroitin sulfates A & C will have marked loss of staining |
what comprises the buffer solution | potassium phosphate diabasic, sodium phosphate dibasic PH of 6 |