Lung cells were dissociated as previously described (Sharma et al., 2022 (link)). The right lung was dissociated to a single cell suspension with Multi Tissue Dissociation Kit 2 (130–110–203, Miltenyi) according to the manufacturer’s protocol using the gentle MACS Dissociator (Miltenyi). The cell suspension was filtered through a 70 μm strainer, pelleted by centrifugation, and cleared of red blood cells with ACK Red Blood Cell Lysis buffer (BP10-548E, Lonza). Live cells were enumerated using trypan blue exclusion counting on a Countess Automated Cell Counter Hemocytometer C10227 (Invitrogen). Lung cells were then used as described below for Fluorescence-Activated Cell Sorting (FACS) analysis. Additionally, ten million lung cells per animal were used for CD45+ enrichment using the CD45 MicroBeads kit (cat# 130–109–682, Miltenyi) per manufacturer’s instructions.
Countess automated cell counter hemocytometer c10227
Manufactured by Thermo Fisher Scientific
The Countess Automated Cell Counter Hemocytometer C10227 is a compact and easy-to-use device designed for automated cell counting. It utilizes digital image processing technology to provide accurate and reproducible cell counts. The instrument is capable of analyzing a variety of cell types, including mammalian, yeast, and bacterial cells.
Automatically generated - may contain errors
Lab products found in correlation
Multi tissue dissociation kit 2, by Miltenyi Biotec (2 mentions)
Cd45 microbeads kit, by Miltenyi Biotec (2 mentions)
Macs dissociator, by Miltenyi Biotec (2 mentions)
Ack red blood cell lysis buffer, by Lonza (2 mentions)
Gentlemacs c tube, by Miltenyi Biotec (2 mentions)
Gentlemacs tissue dissociator, by Miltenyi Biotec (2 mentions)
Debris removal solution, by Miltenyi Biotec (2 mentions)
Tumor dissociation kit, by Miltenyi Biotec (1 mentions)
Xrad 320 kv orthovoltage x ray system, by Precision X-Ray (1 mentions)
Tumor dissociation kit protocol, by Miltenyi Biotec (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
7 protocols using countess automated cell counter hemocytometer c10227
1
Lung Cell Dissociation and CD45+ Enrichment
2
Tumor Dissociation and CD44+ Cell Isolation
3
Lung Cell Isolation and Characterization
4
Allogeneic Bone Marrow Transplant in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three donor adult female WAG/RijCmcr rats (11–12 weeks old) were exposed to 7.75 Gy total body irradiation (TBI) or sham irradiation. On day 30 post-irradiation, donor animals were humanely euthanized and both femurs were carefully excised. Bone marrow from individual femurs was flushed using a 28-gauge syringe into 5 ml of sterile phosphate buffered saline (PBS) containing 10% fetal bovine serum. Cells were pelleted by centrifugation and the red blood cell fraction was removed with (Ammonium Chloride Potassium) ACK lysis buffer. Total viable cells were determined using a Countess Automated Cell Counter Hemocytometer C10227 (Invitrogen) with trypan blue exclusion method. Adult female WAG/RijCmcr rats bone marrow recipients were conditioned with 13 Gy TBI. All rats were irradiated without the use of anesthetics by being placed in a plastic jig and the entire body was exposed using a XRAD 320 kV orthovoltage x-ray system (PrecisionX-Ray, Madison, CT). The X-ray system was operated at 320 kVp and13 mAs with a half-value layer of 1.4 mm copper and a dose rate of 169 cGy/min. Recipient rats received 5 × 106 whole bone marrow cells in a volume of 300 ul via intravenous tail vein injection at 24 h post-irradiation.
5
Rat Bone Marrow Cell Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow was harvested from the femurs of euthanized rats. Each femur was carefully excised from the animal and flushed with PBS containing 10% FBS and 1% Pen/Strep using a 1 ml syringe with a 25G needle, making sure to flush from both ends to ensure all cells were removed. Cells were pelleted by centrifugation and the red blood cell fraction was removed with ACK lysis buffer. Total viable cells were determined using a Countess Automated Cell Counter Hemocytometer C10227 (Invitrogen) with trypan blue exclusion method.
6
Comprehensive Tumor Tissue Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Biopsy specimens were obtained fresh at the time of surgical resection or as part of a research-specific biopsy between February 27, 2022 and March 15, 2023. Samples were washed in PBS with 4% antibiotic-antimycotic (Gibco). The specimen was then split for multiple parallel analytic pathways. A piece was preserved in 10% buffered formalin for subsequent histologic analysis. The remaining tumor was diced into segments and separated for (1) immediate flow cytometry, (2) malignant cell culture, (3) TIL culture. The tumor piece for flow cytometry was then digested according to the Miltenyi Tumor dissociation kit protocol in a Miltenyi gentleMACS C tube using the Miltenyi gentleMACS tissue dissociator. Cellular debris was removed from the dissociated cell suspension using Debris Removal Solution (Miltenyi). Live cells were enumerated using trypan blue dye on Countess Automated Cell Counter Hemocytometer C10227 (Invitrogen).
7
Isolating Rat Bone Marrow Cells
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!