CI-191L Air-Jacketed CO2 Incubator Usage Report（二）

01. Objective

To compare the performance of the CI-191L air-jacketed CO₂ incubator with that of another commercial CO₂ incubator in cultivating different cell types, focusing on cell proliferation and apoptosis.

02. Detection Methods

Proliferation assays: BrdU, CFSE, and eFluor450

Apoptosis detection: Annexin V/7-AAD apoptosis detection kit

In Part 1 of this usage report, we introduced the three methods used for proliferation analysis. In this section, we focus on the apoptosis detection results.

03. Apoptosis Detection

3.1 Annexin V/7-AAD Apoptosis Assay

3.1.1 Principle of the Experiment

In healthy viable cells, phosphatidylserine (PS) is normally located on the inner leaflet of the plasma membrane. However, during early apoptosis, PS translocates from the inner to the outer leaflet, becoming exposed on the cell surface. Annexin V is a 35–36 kDa Ca²⁺-dependent phospholipid-binding protein that binds with high affinity to PS. This property allows Annexin V to serve as a marker for early apoptotic cells by binding to the externalized PS.

7-AAD is a DNA-binding dye that cannot penetrate the intact membranes of viable or early apoptotic cells, but can enter late apoptotic or necrotic cells and bind to their DNA. Therefore, when Annexin V is used in combination with 7-AAD, the following staining patterns are observed:

Annexin V-/7-AAD-: Viable cells

Annexin V+/7-AAD-: Early apoptotic cells

Annexin V+/7-AAD+: Late apoptotic or necrotic cells

3.1.2 Experimental Procedure

1)Collect B cells cultured for 48 hours. Wash the cells with 1 mL of pre-chilled PBS, centrifuge at 3400 rpm for 5 minutes at 4ºC, and discard the supernatant.

2) Wash the cells once with 1 mL of 1× Annexin V Wash Buffer. Centrifuge again at 3400 rpm for 5 minutes at 4ºC and discard the supernatant.

3) Add 60 μL of staining solution to each sample tube, containing APC-Annexin V (1:400 dilution in 1× Annexin V Wash Buffer). Mix gently and incubate at room temperature in the dark for 13 minutes. Then add 5 μL of PerCP/Cy5.5-7AAD to each tube, and incubate in the dark for an additional 5 minutes at room temperature.

4)Resuspend the cells in 200 μL of 1× Annexin V Wash Buffer and transfer the samples to flow cytometry tubes. Analyze the samples using a flow cytometer and visualize the results using FlowJo software.

3.1.3 Experimental Conclusion

Primary mouse B cells were cultured simultaneously in both incubators under identical conditions for two days. Apoptosis was evaluated using flow cytometry. The results showed no statistically significant difference in early or late apoptosis between the two incubators. This indicates that both CO₂ incubators provide comparable and reliable cell culture performance.

 

04. User Experience

1) Highly efficient automatic high-temperature sterilization

This function is convenient and time-saving for researchers. Authoritative testing confirmed a sterilization rate of over 99.999% against the following organisms: Escherichia coli (8099), Staphylococcus aureus (ATCC 6538), Candida albicans (ATCC 10231), Aspergillus niger (ATCC 16404), Aspergillus flavus (AS3.3950), and Trichoderma viride (AS3.2942).

2) Comparable to leading brands in cell culture performance

Based on experimental results, the CI-191L CO₂ incubator meets the requirements for culturing most common cell types.

3) Auto-restart after power failure

The incubator automatically resumes operation after a power outage, reducing the risk of cell death caused by prolonged downtime.

4) Stable and quiet operation with minimal vibration

Ideal even for sensitive cells such as brain and neuronal cells, ensuring high viability and stable growth conditions.

5) Convenient data access and remote monitoring

When connected to a Wi-Fi monitor, users can remotely track real-time data and adjust settings directly from their mobile phones.

6) User-friendly color touchscreen interface

The intuitive design makes operation simple and clear at a glance.