Aim:-To study the preparation of permanent slide by micro technique using microtome.
Requirements:-coupling jar , reagent bottles,slide,coverglass,spirit,puraffin wax,beaker,l-blocks,lamp, microtome,blade,wooden block.
General outline:-
Animal tissue for microtome ->necrotizatio of animals ->fixing of tissues ->washing ->dehydrotion->claring or dealcoholization of tissue ->Embedding of tissue ->block making ->trimming ->section cutting ->double staining, dehydration, clearing and mountings ->microscopic study ->microscopic photography.
Procedure:-
Animals aur tissue for microtone normally frog rat black red are favourable for microsome is in vertebrate animals file in invertebrate earthworm is used.
Necro Jason of animals generally chloroform is used to nephrotic necrotise animal take a big jar and make a cotton bed on bottom of there bottom of jar
Sprinkle the chloroform on the cotton and put animal in the year in the jar
After 15 to 20 times when animal fully entities take out the animal and keep it in desceting frog and asect the animal
Fixing of tissue tissue is generally fixing in the bounds floats C for 100 ml saturated picnic acid 75 ml floram 20 ml glycerine acetic acid 5 ml
Other pics you like acetone formalin methanol also can be used
Washing of tissue after the fixation of 24 hours take out the tissue from the fixit and keep it under slow running tape water for 24 hours
The induction of complete removal of picnic acid comes when no yellow is water is seen
Dehydration for the dehydration of tissue one should have to pass through alcohol series
The alcoholsation or clearing of tissue normally the clearing agents are the xylene and cdwood oil first keep the tissue in solution which content region one alcohol xylem ratio the solution release by one M1 alcohol xylem ratio
Embedding wax is used for embeding tissue take a tissue from xylene and keep it in a bigger containing Island Max 4 30 minutes then transfer that issue in methyl wax for ambouring for 1 to 2 hours
Block making make the block either in the metal elsept angle or in paper boat or cavity before block making apply glycerine on internal surface of the blocks poor the melted wax at the bottom of refrigerator cavity formed by l block at tissue and feather and more melted wax over the tissue to the cover during block making take a care of out of air bubbles
Dreaming dream the wax around the embedded material and the make a permanent triangular block on the one side keep sufficient space of the block of a fixing on the black holder
Section cutting the blocks are cut either by rocking microtome or rotary microsome at the six micro thickness ribbon are Cape over measures arbi mean cottage night keep the slide ready apply pin had had mayor beam in over the right and the rabbit by last finger where in the speaking in the section over glass surface give to three rows of section depending up on the size of the section
Double staining the hydration clearing mounting double staining is applied hemosis and is in nucleus and cytosin of the cell repres in take the individual slide and keep it in dialling to remove x45 to 10 minutes now wax is removed from the xylem tissue is free now.
Aim:-To study the preparation of permanent slide by micro technique using microtome.
Requirements:-coupling jar , reagent bottles,slide,coverglass,spirit,puraffin wax,beaker,l-blocks,lamp, microtome,blade,wooden block.
General outline:-
Animal tissue for microtome ->necrotizatio of animals ->fixing of tissues ->washing ->dehydrotion->claring or dealcoholization of tissue ->Embedding of tissue ->block making ->trimming ->section cutting ->double staining, dehydration, clearing and mountings ->microscopic study ->microscopic photography.
Procedure:-
Animals aur tissue for microtone normally frog rat black red are favourable for microsome is in vertebrate animals file in invertebrate earthworm is used.
Necro Jason of animals generally chloroform is used to nephrotic necrotise animal take a big jar and make a cotton bed on bottom of there bottom of jar
Sprinkle the chloroform on the cotton and put animal in the year in the jar
After 15 to 20 times when animal fully entities take out the animal and keep it in desceting frog and asect the animal
Fixing of tissue tissue is generally fixing in the bounds floats C for 100 ml saturated picnic acid 75 ml floram 20 ml glycerine acetic acid 5 ml
Other pics you like acetone formalin methanol also can be used
Washing of tissue after the fixation of 24 hours take out the tissue from the fixit and keep it under slow running tape water for 24 hours
The induction of complete removal of picnic acid comes when no yellow is water is seen
Dehydration for the dehydration of tissue one should have to pass through alcohol series
The alcoholsation or clearing of tissue normally the clearing agents are the xylene and cdwood oil first keep the tissue in solution which content region one alcohol xylem ratio the solution release by one M1 alcohol xylem ratio
Embedding wax is used for embeding tissue take a tissue from xylene and keep it in a bigger containing Island Max 4 30 minutes then transfer that issue in methyl wax for ambouring for 1 to 2 hours
Block making make the block either in the metal elsept angle or in paper boat or cavity before block making apply glycerine on internal surface of the blocks poor the melted wax at the bottom of refrigerator cavity formed by l block at tissue and feather and more melted wax over the tissue to the cover during block making take a care of out of air bubbles
Dreaming dream the wax around the embedded material and the make a permanent triangular block on the one side keep sufficient space of the block of a fixing on the black holder
Section cutting the blocks are cut either by rocking microtome or rotary microsome at the six micro thickness ribbon are Cape over measures arbi mean cottage night keep the slide ready apply pin had had mayor beam in over the right and the rabbit by last finger where in the speaking in the section over glass surface give to three rows of section depending up on the size of the section
Double staining the hydration clearing mounting double staining is applied hemosis and is in nucleus and cytosin of the cell repres in take the individual slide and keep it in dialling to remove x45 to 10 minutes now wax is removed from the xylem tissue is free now.
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