HP - TEMP & NUMBERS Flashcards
temp of WATER BATH?
45-50°C / 6-10°C LOWER than the MP of wax
temp for Rapid Substitution/Rapid Freezing
-160 to -180°C
temp of Paraffin Oven during INFILTRATION?
2-5°C ABOVE the MP of wax
temp of Paraffin Oven during EMBEDDING?
5-10°C ABOVE the MP of wax
VOL. OF RGTS?
F?
D?
D?
C?
I?
Fixation- 20x
Decal-20x
Dehy-10x
Clearing-10x
Infiltration-25x
Volume of fixative for prolonged fixation
50-100x
vol of fixative for COLON POLYP?
size of colon polyp is
2 x 2 x 3 cm = 12 cm
12 x 20 = 240 ml of FIXATIVE
Water must be removed by heating the wax at what temp?
100-105 deg C
Thickness of tissue section:
Frozen section?
Paraffin section (Rotary/rocking)?
Celloidin section (sliding)?
Semithin?
Ultrathin?
Frozen section = 4 micra
Paraffin section (Rotary/rocking)= 4-6 micra
Celloidin section (sliding) = 10-15 micra
Semithin = 0.5 - 1 micra
Ultrathin = 60-100 nm
Temperature of liquid nitrogen
-170°C (-160 TO -180°c)
MICROTOME KNIFE
One side of the knife is flat/straight while the other is concave
Base-sledge/Rocking/Rotary microtome
MEASUREMENT?
PLANE CONCAVE
(25mm in length)
MICROTOME KNIFE
with both sides concave (ROUTINE PARAFFIN SECTIONS)
Recommended for ROTARY MICROTOME
MEASUREMENT?
BICONCAVE
(120mm in length)
MICROTOME KNIFE
have both sides straight (FROZEN SECTIONS and extremely HARD AND TOUGH specimens)
Base-sledge/Sliding microtome
MEASUREMENT?
PLANE WEDGE
(100 mm in length)
Most routine cryostat work is performed with chamber temperature of
commonly -18 to -20°C (ARC: -20°C)
Refractive index of glass
1.518
traditional temp of HP?
22-25°C
temp of Autotechnicon?
40°C
✓ Fixation temperature:
✓ Normally used:
✓ Fixation temperature: RT (18-30°C)
✓ Normally used: 20-22°C
- Fixative Osmolality
- Hypertonic solutions can cause what?
- Isotonic (340 mOsm)/Hypotonic solutions cause what?
- Added to Osmium tetroxide fixatives for EM:
- slightly hypertonic solutions (400-450 mOsm)
- shrinkage
- swelling and poor fixation
- Sucrose
- Fixative pH:
- The Optimum temperature range for fixation is:
- It accelerates fixation but hastens autolytic changes and enzyme destruction
- Fixative pH: Neutral (6.0-8.0)
- 37-56°C
- Moderate Heat
temp:
- Tissue processors:
- Electron Microscopy and Histochemistry:
- Mast cells for EM:
- 40-42°C
- 0-4°C
- RT
Nucleic acids fixation temp?
Rapid at higher temp
temp:
Rapid fixation of very urgent biopsy specimens-
Formalin heated to 60°C
temp:
Fixing tissues with tuberculosis-
Formalin heated to 100°C
- Dehydration is accomplished through the use of:
- Starting Concentration:
- Final Concentration:
- increasing grades of alcohol
- Starting Concentration: 70%
- Final Concentration: 100%
- Infiltration in overheated paraffin above ____°C produces shrinkage and hardening of tissues
- Ideal MP of infiltrating agent:
- Normally used for routine work:
- 60°C
- 56-58°C
- 56°C (56-60°C)
The knife is usually tilted at ____° angulation on a microtome to allow a clearance angle between the cutting facet and the tissue block
0-15
When specimen is more than a few drops, the fluid should be centrifuged first. This is usually done at moderate speed at _____ RPM for ___ minute(s).
2000 RPM, 2 minutes
By far the best method to collect cells from body fluid (e.g., urine, pleural or peritoneal fluid):
✓ Cytospin Preparation (1000 RPM for 1 minute)
vacuum embedding involves wax impregnation under _____ atmospheric pressure. A vacuum pressure of up to ____ is applied using a water or mechanical pump
Negative pressure; 760 mmHg
recommended temp of decalcification?
18-30 C
Melting point of the embedding medium that is used as a substitute for paraffin wax and is a mixture of highly purified paraffin wax and synthetic plastic polymers.
56-57 C
An angle refers to the angle formed between actual cutting edges.?
Bevel angle (27-32 deg)
This is an angle which ensures maximum penetration of tissues and minimizing distortion.
Inclination angle (15°)
