Subculture of a Cell Line

As cultured cells grow and divide, they use up nutrients and eventually, cells will multiply to a point that they run out of space in the culture vessel.
Subculturing is the process of diluting and transferring the cells into more vessels with fresh culture medium so we can produce a new culture and the cells can continue growing.
In this video, we demonstrate the steps for propagating a culture by transferring NIH3T3 cells from a previous culture to a new culture vessel.
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[@ShakespeareCafe]

Invest in autoclaved glass pipettes and a vacuum pump and save money and plastic waste using so many serological pipettes. Also, aliquot your media into 50 ml falcon centrifuge tubes to prevent contamination of the large media bottle; sharing media is the number 1 source of contamination. Media is expensive.

[@donnaandre7633]

What a beautiful lab!

[@NCBioNetwork]

Glad you like. Yes, it is a nice lab!

[@user-mm8cq9xg3m]

Best vedio. Thank you very much

[@user-bd1tb5zr2z]

시청완료!

[@Tv7_telugu]

Awesome

[@drpujaprasad6889]

Hi I want to setup a lab like this. How much does fully equipped lab like this would cost to setup ?

[@rasoihargharki342]

Nice

[@funny11744]

How to prepare a trypan solution 0.4% starting from powder 60% trypan blue ?

[@kosheeka]

To prepare a 0.4% trypan blue solution from 60% trypan blue powder, follow these steps: Calculate the weight of 60% trypan blue powder needed: • Use the equation Concentration 1 × Weight 1= Concentration 2 × Weight 2 60 % × Weight of powder = 0.4 % × 100 mL Solving this, Weight of powder = 0.4 × 100 ÷ 60 = 0.667 grams • Add 0.667 grams of 60% trypan blue powder to approximately 90 mL of distilled water. 9 Stir until completely dissolved. • Make up the final volume to 100 mL with distilled water. This will yield a 0.4% trypan blue solution.

[@funny11744]

@@kosheeka Thank you

[@binazubayraeva5710]

how can I count confluency?

[@JNNT934]

Bina Zubayraeva If you need an acculturation of cell confluence, you may use the ImageJ program to measure the cell confluence. However, when you working in culture room to maintain cell culture, you should be estimated by your eyes. It will be faster to work.

[@kosheeka]

Visual Estimation • Low confluency: Sparsely distributed cells. • Medium confluency: Cells are visible but with clear spaces between them. • High confluency: Cells form a nearly complete monolayer with minimal empty spaces. • Confluent: Cells form a complete monolayer, often with overlapping cells. Manual Cell Counting with Hemocytometer • Prepare a cell suspension. • Load the hemocytometer. • Count cells in specific grid squares. • Calculate cell concentration. • Estimate confluency based on cell density and culture vessel area. Automated Image Analysis • Capture images of the cell culture. • Use image analysis software (ImageJ) to identify and count cells. • Calculate confluency based on the cell-covered area.

[@patisserie1]

What are the qualifications needed to do this kind of Lab work.

[@drpujaprasad6889]

I guess too many qualification are not required for doing this kind of lab work. 10+2 school study u could do ur bachelors then marsters and Phd. or u could just be a lab tech. The lab tech job doesn't require much qualifications.......graduation in cllg and then 1 to 2 years traning as lab tech.

[@Bm-hm7tt]

Really difficult field to get into. Graduated over a year ago and still trying🤷🏻‍♀️. This is my teacher though so ayee

[@Ankitsingh-qf6el]

I am doing this in my bachelor's

[@scholars789]

well ..

[@weimullerjohann9118]

Microscope is to cheap ! Cell cultures are placed in the incubator and then placed under the microscope. You don't give anything to gangsters.