High Efficiency Particulate Air Filter HEPA
CO 2
74 30 min 2 min
CO 2 37 5 CO 2 CO 2 90 14 h) 3000
CO 2 4-20 -80-196 1000 2500 rpm Zeiss
70
1. 5 2. 3. 4. 15 20
1 2 3 4 2 NaOH 5 6 5 30 min 7
160~180 2 h 120 20 min Zeiss 75 1
100 100
BSS ph EDTA
a b c
TC199 MEM RPMI-1640 DMEM
5 10
Eagle Eagle Eagle s minimum essential medium MEM BEM Basal Eagle Medium DMEM Dulbecco MEM DMEM 4500 mg/l 1000 mg/l
RPMI 1640
Ham Ham F7 F10 Ham F12
NaHCO 3 100 U/ml 100U/ml ph 7.0~7.2 PH 0.2 0.22 m+0.45 m 4 10%
Serum free medium, SFM DNA
SFM
1975 Sato 20
56 30
1. 2. A 3. 0.1~0.5%
80 95 5 20 2.0 g/l 100
RPMI-1640 1 2.0 g 100 / 1000 ml, ph 7.2 10
1 2 3 RPMI-1640 4
0.2% 30 50 min 75 30 min 75%
30 min 75
3T3-L1 3T3-L1
MG-63 MG-63
3 1 1000 rpm/min 2 3 0.25
1. 2. 1 2 ml 0.25 2 10 min 3. 4. 5. 6. 1/10 1/40 7. 8.
Coulter
1 2 3 3 4 /ml 4 / 4 10000 10%
Preparation of microscope/hemocytometer for cell counting. Position cover slip so that glass covers both counting areas equally (See figure below). Filling the hemocytometer chamber (Left).
Coulter = 4 x 10 4 x 4 = /ml
Growth of yeast at ph=7 (Number of cells count/ml vs.time) Number of cells count/ml 1.00E+09 1.00E+08 1.00E+07 3667778 4485556 49111111 1.27E+08 Control 1.00E+06 0 5 10 15 20 25 30 Time (hr)
1 1 0.5 ml 2 0.5 ml 0.4% 2 3 min 3 4
% = / x 100%
2 MTT MTT MTT formazan) DMSO formazan OD MTT
MTT 1 96 10 3-10 4 / 200 96 370 37 5 CO 2 2 2 MTT 50 / 3 3 DMSO 150 / 10 4 OD 570 nm
-25 1 2 /min -25 5 10 /min -100 1 2 40 30
DMSO
1. 20% 10% DMSO DMSO 2., 1 10 6 5 10 6 /ml) 3. 1 ml
-25 1 2 /min -25 5 10 /min -100 4 10 --->-20 30 --->- 80 16 18 ( )--->
1. 37 38 1 min 2. 5 min 10 3. 1200 rpm 4 min 4.
1. 0.1 0.8 µm 0.22 0.45 µm 2. 3. 4. 5. 6. 7.
Chlamydia pneumoniae associated with macrophages (RAW cells). A. Typical pear shaped elementary bodies (EBs) (arrows) are shown at the macrophage surface. B. Intracellular EBs.
PCR Hayflick
Hayflick 3 5 ( M. hyorhinis)
bisbenzimide, Hoechst 33258 DNA Adenosine-Thymidine (A-T) rich DNA A-T 55 80%
PCR 16s 23s 1 5 - -3 1 5 - -3 2 5 - -3 2 5 - -3 PCR
:
1. 5 10 24 48 2. 41 5 10 3.
G+ 100 IU/ml G- 100 µg/ml G+, G- 200 µg/ml G+, G- 10 µg/ml G+, G- 50 µg/ml 2 µg/ml 25 µg/ml
PCR