Morphological observation experiments on cultured cells
Morphological observation experiments on cultured cells
The Morphological Observation of Cultured Cells experiment is an experiment in which the morphology of cells is observed through a microscope to make a determination of their growth.
Principle
The basic principle of the morphology observation experiment of cultured cells is that the growth state of cultured cells can be determined by observing the number of particles, the transparency and the clarity of the outline of the cultured cells under an inverted microscope.
Live cells in good growth state, its cytoplasm is usually transparent, homogeneous, cytoplasmic particles are less, the cell outline is very shallow, not obvious. With the prolongation of the culture time, the granular material in the cell gradually increases, the transparency of the cell is weakened, the outline of the cell is enhanced, and the number of nucleoli also increases.
Operation method
Morphological observation experiments on cultured cells
Principle
The basic principle of the morphology observation experiment of cultured cells is that the growth state of cultured cells can be determined by observing the number of particles, the level of transparency and the clarity of the outline of the cultured cells under an inverted microscope. Live cells in good growth state, its cytoplasm is usually transparent, homogeneous, cytoplasmic particles are less, the cell outline is very shallow, not obvious. With the prolongation of the culture time, the granular material in the cell gradually increases, the transparency of the cell is weakened, the outline of the cell is enhanced, and the number of nucleoli also increases.
Materials and Instruments
Equipment: Move The basic procedure of morphological observation of cultured cells can be divided into the following steps: Caveat 1. When observing cell morphology, aseptic operation is required to avoid contamination. 2. In order to reduce the influence of the external environment on the growth of cells, the observation of cells should not be too long, and when observing cells in multiple vials, they should be taken and placed in batches. For more product details, please visit Aladdin Scientific website.
① Hepatocytes in subculture for 3 days
② Hepatocytes in subculture for 7 days
(iii) Dermal fibroblasts in subculture for 3 d.
④ Dermal fibroblasts cultured for 7 d. ⑤ Inverted microscope.
⑤ Inverted microscope
⑤ Inverted microscope ⑥ Ultra-clean bench
⑦ CO
2
Incubator
Reagents: ① D-Hanks liquid
① D-Hanks liquid
② DMEM medium (containing 10% calf serum)
(1) Observation of the morphology of cultured cells
A. Place the cultured hepatic cells for 3 d under an inverted phase contrast microscope and observe the morphology of the cultured cells, the arrangement of the cells, and the degree of connectivity between the cells.
B. Record the results of the observation.
C. Place the hepatic cells back into the incubator, and take out the cultured hepatic cell from the cultured fibroblast for 3 d.
D. Observe and record the results of the morphology, cell arrangement, and degree of connectivity of the fibroblasts with the inverted phase contrast microscope. D. Observe the morphology of the fibroblasts, the arrangement of the cells and the degree of connection between them with an inverted phase contrast microscope and record the results.
E. Compare the recorded results of the morphological characteristics of the hepatocytes with those of the fibroblasts.
(2) Observation of cell morphology, structure and growth
A. Place hepatocytes or fibroblasts cultured for 3 d under the 10x objective of an inverted phase contrast microscope.
B. Observe the internal structure of the cells, their transparency, and their contours.
C. Use a 40x high magnification microscope to observe the structure of the cells and their contents.
D. Record the results of the observation and return the cells to the incubator, and at the same time, take out the hepatocytes or fibroblasts that have been cultured for 7 days. Hepatocytes or fibroblasts from 7 d of passaged culture were placed under an inverted phase contrast microscope.
E. Observe the transparency, cell outline and contents of the cells with 10x and 40x objective lenses, respectively.
F. Record the results, and compare with that of the cells cultured for 3 d of passaged culture.