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Section D
Live Dye Partial Organells and Observe Them with Microscope
April,2013
Department of Cell Biology School of Basic Medical Sciences
Xinjiang Medical University
Nafeisha Kadeer
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Purposes:
• Master the method to live dye partial organells--mitochondria and vacuole system.
• Master the basic structure and distribution of mitochondria, vacuole system in cell.
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Contents:
1.Live dye the mitochondria in the rabbit liver cell.
2.Live dye the vacuole system in the swordlike cartilage cell of toad.
3.Observe the Golgi Apparatus.
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Materials
1. microscope,slide,coverslip,pipette, tweezers,absorption tissue,dissection
plate,dissection needle,dissection scissors,eye scissors,culture vessel,nails,thick plastic gloves;
2. a rabbit,a toad;3. 1/300Jaun’s Green 1/3000Neutral red 0.85% Ringer solution 0.65% Ringer solution
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1 、 The method to live dye the mitochondria in the rabbit liver cells
Principle
Jaun’s Green is a special live dye for mt. The
cytochrome oxidase in the mt can oxidize this
dye into blue particles, so we can examine the
existence of the mt by these blue particles.
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Inject air into rabbit’s ear vein
vein
artery
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Method • Kill a rabbit (inject air into it’s ear vein) , put it on
the dissection plate on back, then open it’s abdominal
cavity. Cut down a small piece of liver tissue from thinnest rim, put it into cultured dish, with Ringer’s solution wash off the blood.
• Transfer liver tissue on slide, make it vertial. From it’s bottom add one drop of Jaun’s Green to half-cover it, stained for about 20 minutes.
• Dispart tissue using tweezer, remove the large blocks tissue. Finally some single cells are left on slide.
• Add a drop of Ringer’s solution, then cover it with coverslip, absorb excess solution.
• Observe your sample with microscope.
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Observe the slide in the microscope.
Using the objective of 10x magnification, then
the objective of 40x magnification.
We can see there are many blue particles in the
cytoplasm of the liver cells, which are arranged
tightly. These blue particles show location of mt.
Result:
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Mitochondria in liver cell
nucleus
cell membrane
mitochondria
(high power objective)
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2 、 How to live dye the vacuole system in the swordlike cartilage cells of toad
Principle
In cytoplasm of animal cell, the vesicles enclosed by the
membrane all belong to vacuole system (except mt), including
Golgi Apparatus 、 Lysosome 、 Endoplasmic
Reticulum , etc. The cartilage cells contain abundant vacuole
system. However, these vacuoles are too small to be seen if they
are not stained with dye. Neutral red can specially stain the
vacuole system dark red and stain cytoplasm and nucleus light
red. So we can observe location of vacuole system .
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Method • Kill a toad, open it’s abdominal cavity, then expose
the swordlike cartilage of breastbone.
Cut down a small piece of cartilage tissue from its thinnest rim, put it on slide.
• Add a drop of Neutral red to dye for 15 minutes (not more than 15 min).
• Remove the dye using the absorbent tissue. Add a
drop of Ringer’s solution, add coverslip.Absorb
excess solution.
• Observe your sample with microscope.
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Swordlike cartilage of toad
swordlike cartilage
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Under microscope, we see cartilage cell is
oval. There are many rosy vesicles .These
rosy vesicles are the vacuole system.
Result:
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3 、 Observe Golgi Apparatus
There are prepared slide of rabbit nerve
knot. First observe it using low power. We can
see many oval neurons. It is oval and stained yellow. Then observe
the neurons using high power.
We can see the round and transparent central parts are nucleus of
neuron. In cytoplasm there are many dotlike 、 sticklike and
threadlike structures scattered and stained dark tan. These dark tan
structures are Golgi Apparatus.
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(high power objective)
nucleus
nucleolus
Golgi Apparatus