cell-culture(细胞培养前准备)

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Cell culture aseptic technique

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Basic knowledge

How to design Cell Culture Room ? What general apparatus we need ? How to prepare for cell culture ?

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

1. Preparing room 2. liquor room 3. Cell culture room

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Cell culture room

centrifuge PC gate super Clean bench gate Preparing room

Dressing zone

Microscope

corridor

gate

cabinet Refrigerator

incubator

Sterile zone

Buffer zone

gate Liquor room

Gate

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Super clean bench Bacterial free zone

Dressing zone

Buffer zone

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Preparing room

Distilled water

Autoclave sterilizer

drying and baking Bench for package

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Liquor room

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Treatment of glassware

flask

bottle

pipette

dish

tube

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

New glassware treatment

1. Scrubbing by tap water Purpose: get rid of dust 2. To Immerge by the dilute hydrochloric acid for 12 hours after drying .

Purpose: get rid of metals such as lead and arsenic

3. Scrubbing by tap water and drying by baker,

To immerge by concentrated sulfuric acid 24 hours.

Purpose: get rid of all impurity

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

4. Scrubbing by tap water by distilled water

15 times 10 times

by double distilled water 3 times 5. Drying by baker

6. Sterilizing by autoclave after packaging

Used glassware treatment

Step3 step 6

IF we have a lot of fund support, we can use disposable material.

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

summary

Acid treatment (new or used) Scrubbing (tap or distilled water) Sterilizing( autoclave, alcohol or ultraviolet)

tip

transferpettor

Multi-well plate

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Aseptic technique

Laboratory : 1. Cleaning the floor and surface of apparatus by benzalkonium bromide per week . 2. Sterilizing by ultraviolet before experiment . Human : 1. Cleaning hand and dress the aseptic clothes, hat ,mouth-muffle and slipper. 2. Sterilizing hand by 75% alcohol before operation .

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Cell culture protocol (digest by enzyme )

1. Preheating medium at 37 ℃. 2. Washing cell 2 times by Hanks or PBS. 3. Abandoning liquid and adding trypsin . 4. Observing cell in microscope. 5. Replacing fresh medium to cease digestion. 6. Making cell suspension and cell counting. 7. Inoculating the cell to a new flask or dishes. 8. Incubating in 37 ℃, 5%CO2 environment.

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Hanks:

salt solution

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

Super clean bench or Bio- safety Cabinet

普通细胞培养室的布局;培养器皿的使用原则;一般细胞传代培养的流程。全部图片化展示。

CO2 cell incubator