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Automated counting system

Manufactured by Cellular Technology

The Automated Counting System is a laboratory equipment designed to perform automated cell counting and analysis. The core function of this device is to accurately enumerate and quantify various cell types in a sample. The system utilizes advanced imaging and analysis algorithms to provide precise cell counts, viability assessments, and other relevant cell population metrics.

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2 protocols using automated counting system

1

Neutralizing Antibody Assay for HCVcc 1a/2a Chimera

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HCVcc 1a(H77)/2a chimeric virus was generated as previously described.36 The E1E2 region was based on the H77 consensus sequence (accession no.: AF009606). This region (aa 192‐746 of the HCV polyprotein) shares 94.8% homology with the genotype 1a E1E2 sequence used for the phase I vaccine study (HCV‐1; accession no.: M62321; http://onlinelibrary.wiley.com/doi/10.1002/hep.28108/suppinfo). Neutralizing antibody titers were determined using Huh7.5 cells by performing 2‐fold dilutions of sera from 1:32 to 1:1,024 and mixing 1:1 with HCVcc 1a/2a (100 focus forming units [ffu]/50 μL), as previously described,36 to give starting dilutions of 1:64. After 72 hours, cells were fixed (2% formaldehyde), stained with anti‐HCV core mouse mAb (6G7; diluted 1:1,000), and developed with the Diaminobenzidine‐staining ABC Kit (Vector Laboratories, Burlingame, CA), as previously described.37 Foci were counted using an automated counting system (Cellular Technology Limited [Cleveland, OH] using BioSpot 5.0 software).
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2

Generation and Neutralization of HCV Chimeric Virus

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HCVcc 1a(H77)/2a chimeric virus was generated as previously described (36 (link)). The E1E2 region was based on the H77 consensus sequence (Accession number AF009606). This region (aa 192–746 of the HCV polyprotein) shares 94.8% homology with the Genotype 1a E1E2 sequence used for the Phase I vaccine study (HCV-1, Accession number M62321) (Supplemental Figure S1). Neutralizing antibody titers were determined using Huh7.5 cells by performing 2-fold dilutions of sera from 1:32 to 1:1024 and mixing 1:1 with HCVcc 1a/2a (100 ffu/50μl) as previously described (36 (link)) to give starting dilutions of 1:64. After 72 hours cells were fixed (2% formaldehyde), stained with anti-HCV core mouse monoclonal antibody (6G7) (diluted 1:1000) and developed with Diaminobenzidine-staining ABCKit (Vector Laboratories, Burlingame, CA) as previously described (37 (link)). Foci were counted using an automated counting system (Cellular Technology Limited using BioSpot 5.0 software).
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