To evaluate microglia cell death, we used phycoerythrin-conjugated annexin V (V-PE) and 7-amino-actinomycin D (7-AAD; Guava Nexin® Reagent, #4500-0450, Merck Millipore, Billerica, MA, USA) to determine the percentage of viable, early-apoptotic and late-apoptotic/necrotic cells by flow cytometry. After incubation, plated microglia were trypsinized and added to cells in the incubation media, which were then stained with annexin V-PE and 7-AAD, following manufacturer’s instructions, and analyzed on a Guava easyCyte 5HT flow cytometer (Guava Nexin® Software module, Millipore), as previously described (Barateiro et al., 2012 (link)). The three populations of cells that can be distinguished by this assay are the viable cells (annexin V-PE and 7-AAD negative), the early apoptotic cells (annexin V-PE positive and 7-AAD negative), and the cells in late stages of apoptosis or dead cells (annexin V-PE and 7-AAD positive).
Guava nexin software module
Manufactured by Merck Group
The Guava Nexin® Software module is a component of the Guava flow cytometry platform by Merck Group. It provides core functionality for the acquisition and analysis of cell-based assays.
Automatically generated - may contain errors
6 protocols using guava nexin software module
1
Evaluating Microglia Cell Death
2
Annexin V Assay for Apoptosis
3
Annexin V Assay for Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phosphatidylserine externalization was detected using an Annexin V staining kit (Millipore) following the manufacturer's instructions. Annexin V–positive cells were detected using the Guava System and analyzed with the Guava Nexin software Module (Millipore).
4
Annexin V-based Apoptosis Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phosphatidylserine externalization was detected using an Annexin V staining kit (Millipore) following the manufacturer’s instructions. Annexin V-positive cells were detected using the Guava System and analyzed with the Guava Nexin Software Module (Millipore).
5
Annexin V and 7-AAD for Apoptosis Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phycoerythrin-conjugated Annexin V (Annexin V-PE) and 7-amino-actinomycin D (7-AAD; Guava Nexin® Reagent, no. 4500-0450, Millipore) were used to determine the percentage of viable (Annexin V-PE and 7-AAD negative), early apoptotic (Annexin V-PE positive and 7-AAD negative), and late apoptotic/necrotic (Annexin V-PE and 7-AAD positive) cells by flow cytometry, as described (Gomes et al., 2020 (link)). After incubation, cells were trypsinized and added to the cells already detached in the culture medium. After centrifugation, the pellet of cells was resuspended in PBS containing 1% bovine serum albumin. Then, they were stained with Guava Nexin Reagent according to manufacturer’s instructions and analyzed on a Guava easyCyte 5HT flow cytometer (Guava Nexin® Software module, Millipore).
6
Apoptosis Assessment via Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phycoerythrin-conjugated annexin V (V-PE) and 7-aminoactinomycin-D (7-AAD) mixture (Guava Nexin® Reagent, #4500-0450, Millipore, Billerica, MA, USA) were used to determine the percentage of viable, early-apoptotic, and late-apoptotic/necrotic cells by flow cytometry. After incubation, adherent microglia were collected by trypsinization and added to the cells present in the incubation media. After centrifugation, the pellet of cells was resuspended in PBS containing 1% of bovine serum albumin (BSA), stained with Guava Nexin Reagent according to manufacturer's instruction, and analyzed on a Guava easyCyte 5HT flow cytometer (Guava Nexin Software module, Millipore), as usual in our lab [34 (link)]. Two readings were performed for each sample.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!