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Renal glomerular surface Sample preparation conditions (4 ° C): After fixation with 2% glutaraldehyde perfusion, chopped (thickness approx. 0.5 mm) and re-fixed ⇒ 1% osmium tetroxide fixed 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannin Acid aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 1.5 hour immersion ⇒ Blower, cold air dryer drying Observation Conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
Rat kidney kidney glomerulus Sample preparation conditions (4 ° C): After fixation with 2% glutaraldehyde perfusion, chopped (thickness approx. 0.5 mm) and re-fixed ⇒ 1% osmium tetroxide fixed 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannin Acid aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 1.5 hour immersion ⇒ Blower, cold air dryer drying Observation Conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
Rat kidney proximal tubule (brush border) Sample preparation conditions (4 ° C): After fixation with 2% glutaraldehyde perfusion, chopped (thickness approx. 0.5 mm) and re-fixed ⇒ 1% osmium tetroxide fixed 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannin Acid aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 1.5 hour immersion ⇒ Blower, cold air dryer drying Observation Conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
Rat kidney rlomerular cleavage image (inner blood vessel) Sample preparation conditions (4 ° C): After fixation with 2% glutaraldehyde perfusion, chopped (thickness approx. 0.5 mm) and re-fixed ⇒ 1% osmium tetroxide fixed 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannin Acid aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 1.5 hour immersion ⇒ Blower, cold air dryer drying Observation Conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
Rat kidney proximal tubule (brush border) Sample preparation conditions (4 ° C): After fixation with 2% glutaraldehyde perfusion, chopped (thickness approx. 0.5 mm) and re-fixed ⇒ 1% osmium tetroxide fixed 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannin Acid aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 1.5 hour immersion ⇒ Blower, cold air dryer drying Observation Conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
at liver capillary bile duct Sample preparation conditions (4 ° C): After fixation with 2% glutaraldehyde perfusion, chopped (thickness approx. 0.5 mm) and re-fixed ⇒ 1% osmium tetroxide fixed 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannin Acid aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 1.5 hour immersion ⇒ Blower, cold air dryer drying Observation Conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
Rat liver rascular endothelial cells Sample preparation conditions (4 ° C): After fixation with 2% glutaraldehyde perfusion, chopped (thickness approx. 0.5 mm) and re-fixed ⇒ 1% osmium tetroxide fixed 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannin Acid aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 1.5 hour immersion ⇒ Blower, cold air dryer drying Observation Conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
Rat small intestine Microvilli Sample preparation conditions (4 ° C): After fixing with 2% glutaraldehyde, shred (thickness approx. 0.5 mm) and re-fix ⇒ Fix with 1% osmium tetroxide 2 hours ⇒ DW wash 10 minutes, 6 times ⇒ 1% tannic acid Aqueous solution 3 hours ⇒ DW washing 10 minutes, 6 times ⇒ 1% osmium tetroxide fixation 2 hours ⇒ DW washing 10 minutes, 3 times ⇒ 2% BEL-1 (70% ethanol) 2 hours immersion ⇒ Blower, cold air dryer drying Observation conditions: SE detector (mixed mix), acceleration voltage 3KV, WD20, emission current (0.8 to 4.7μA) |
Rat kidney glomerular cleavage image (inner blood vessel) Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Proximal tubule (brush border) Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Kidney glomerulus Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Rat kidney proximal tubule (brush border) Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Renal glomerular surface Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Rat small intestine soft projection Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Rat liver capillary bile duct Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Rat liver capillary bile duct Double fixing ⇒ 5% BEL-1 (70% ethanol) immersion for 2 hours ⇒ Drying (blower, cold air dryer) |
Neutrophil 1% GA fixation (slide glass with PLL coating) 30 minutes adhesion (wet) ⇒ DW cleaning ⇒ 10% BEL-1 (diluted solution: 100% ethanol) 10 minutes immersion ⇒ Drying (blower → cold air dryer about 15 minutes) Observation conditions : SE detector Upper emission current 5.5μA |
Diatoms 2% GA fixed ⇒ DW cleaning ⇒ 10% BEL-1 (70% ethanol) 10 minute immersion ⇒ Drying (blower, cold air dryer) Observation conditions: Acceleration voltage 3KV Emission current: 5.5μA SE detector: Mix |
Diatoms 1% GA ⇒ DW washing ⇒ 10% BEL-1 (70% ethanol) 10 minutes ⇒ Dry (cold air dryer) |
Diatoms 1% GA ⇒ DW washing ⇒ 10% BEL-1 (70% ethanol) 10 minutes ⇒ Dry (cold air dryer) |
Megalopa larva 1% GA ⇒ DW washing ⇒ 10% BEL-1 (70% ethanol) 10 minutes ⇒ Dry (cold air dryer) |
Megalopa larva 1% GA ⇒ DW washing ⇒ 10% BEL-1 (70% ethanol) 10 minutes ⇒ Dry (cold air dryer) |
Red blood cells 10% (diluted solution: 100% ethanol) 10 minutes immersion Acceleration voltage: 3kV Emission current: 5.5μA SE detector: Upper |
White blood cells 10% (diluted solution: 100% ethanol) 10 minutes immersion Acceleration voltage: 3kV Emission current: 5.5μA SE detector: Upper |
SEM slide glass used 1% GA fixation (SEM slide glass) 30 minutes adhesion (wet) ⇒ DW cleaning ⇒ 10% BEL-1 (diluted solution: 70% ethanol) 10 minutes immersion ⇒ Drying (blower → cold air dryer about 15 minutes) Observation conditions: SE detector Upper emission current 5.5μA |
SEM slide glass used 1% GA fixation (SEM slide glass) 30 minutes adhesion (wet) ⇒ DW cleaning ⇒ 10% BEL-1 (diluted solution: 70% ethanol) 10 minutes immersion ⇒ Drying (blower → cold air dryer about 15 minutes) Observation conditions: SE detector Upper emission current 5.5μA |
SEM slide glass used 1% GA fixation (SEM slide glass) 30 minutes adhesion (wet) ⇒ DW cleaning ⇒ 10% BEL-1 (diluted solution: 70% ethanol) 10 minutes immersion ⇒ Drying (blower → cold air dryer about 15 minutes) Observation conditions: SE detector Upper emission current 5.5μA |
SEM slide glass used < 1% GA fixation (SEM slide glass) 30 minutes adhesion (wet) ⇒ DW cleaning ⇒ 10% BEL-1 (diluted solution: 70% ethanol) 10 minutes immersion ⇒ Drying (blower → cold air dryer about 15 minutes) Observation conditions: SE detector Upper emission current 5.5μA |
Courtesy of St. Marianna University School of Medicine, Electron Microscope Research Facility Chisako Sasaki
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