Estrous cycle stage was determined by vaginal cytology in female mice immediately following testing. Vaginal cytology was assessed by dipping a sterile swab in water and gently swabbing the outer half of the vaginal canal. Vaginal samples were transferred to a microscope slide, air dried, stained using a Hema 3 staining kit (Fisher Scientific, Hampton, NH, USA), dehydrated, and then cover slipped prior to visualization on a light microscope at 20x magnification. Estrous cycle stage was assessed using previously established criteria (Aliagas et al., 2010 (link); Bath et al., 2012 (link)) and were carried out by an observer blind to rearing condition and behavioral results.
Hema 3 staining kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Hema 3 staining kit is a laboratory equipment product designed for the purpose of performing blood cell staining. It provides the necessary reagents and protocols to stain and differentiate various types of blood cells, such as red blood cells, white blood cells, and platelets. The core function of the Hema 3 staining kit is to facilitate the visual identification and analysis of blood cell morphology and composition.
Automatically generated - may contain errors
Lab products found in correlation
24 wellpolycarbonate membrane, by Corning (3 mentions)
Ds ri2 camera, by National Instruments (3 mentions)
Boyden chamber, by Neuro Probe (3 mentions)
Mexicanin 1, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Shandon cytospin 3, by Thermo Fisher Scientific (2 mentions)
Vanox ahbt3 microscope, by Olympus (2 mentions)
Transwell matrigel invasion chambers, by BD (2 mentions)
Eclipse te2000, by Nikon (2 mentions)
Eclipse e200 microscopy, by Nikon (2 mentions)
Transwell insert, by BD (2 mentions)
Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions)
Collagen type 1 gel, by BD (1 mentions)
Matrigel, by BD (1 mentions)
Fluostar optima microplate reader, by BMG Labtech (1 mentions)
Non coated membrane, by Merck Group (1 mentions)
Biocoat matrigel invasion chamber, by Corning (1 mentions)
Biocoat matrigel invasion chamber, by BD (1 mentions)
Te2000 u microscope, by Nikon (1 mentions)
Evos fl auto imaging system, by Thermo Fisher Scientific (1 mentions)
39 protocols using hema 3 staining kit
1
Vaginal Cytology for Murine Estrous Staging
2
Cell Colony Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded into the 6‐well plates at 500–1000 cells per well. The formed cell colonies were fixed with 4% paraformaldehyde (Fisher Scientific) at room temperature for 20 min and stained using Hema 3 staining kit as per the manufacturer's instructions (Fisher Scientific). The number of colonies and the colony area was calculated using imagej (NIH, Bethesda, MD, USA).
3
Fibrocyte Generation with IL-13 and M-CSF
4
3D Cell Migration and Invasion Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For 3D cell migration assays, 105 cells were embedded in 20 μL of type I collagen gel (2.0 mg/mL) extracted from rat tail (BD Biosciences). After gelling, the plug was embedded in a cell-free collagen gel (2.0 mg/mL) within a 24-well plate. After allowing the surrounding collagen matrix to gel (1 h at 37 °C), 0.5 mL of culture medium was added on the top of the gel and cultured for another 2 d. Invasion distance from the inner collagen plug into the outer collagen gel was quantified. For invasion assays, Transwell cell invasion assays were performed using either 24-well polycarbonate membrane (Corning) with 8-μm pore size, or 24-well FluoroBlok Transwell insert (BD) with 8-μm pore size. Inserts were prepared by coating the upper surface with 1 mg/mL Matrigel (BD Biosciences) for 4–6 h at 37 °C in a 5% CO2 incubator. BT549 or 4T1 cells (5 × 104) in DMEM containing 1% FBS were seeded into the upper chamber of the insert. The bottom chamber contained DMEM with 10% FBS. After 24 or 48 h, membranes were processed. Polycarbonate membranes were stained with HEMA3 staining kit (Fisher) and then mounted and enumerated based on number of cells per 20× high power field, five fields per insert. For FluoroBlok transwells, luminescence intensity was measured using a FluoStar Optima microplate reader (BMG Labtech) for 10 individual fields on the bottom of each insert.
5
BAL Cell Isolation and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BAL samples were subjected to centrifugation and cell pellet resuspended in 1 ml PBS. Total cell count in BAL samples was determined by hemocytometer and data expressed as cells/ml. The BAL cells after centrifugation were stained with Hema-3 staining kit (Fisher Scientific) and differential cell count was determined by brightfield microscope.
6
Cell Migration Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For migration assays, cells were harvested and suspended in serum free medium supplemented with 1% BSA. Cell suspension was loaded into the top chamber with a non-coated membrane (Millipore, MA, USA) at a concentration of 1×105 cells per 100 μL. Medium containing 20% FBS was used as a chemoattractant in the lower chamber. After 24 h of incubation at 37°C, cells on the upper surface of the membranes were removed with a cotton swab. The membranes were then stained (Hema3 staining kit; Fisher), and the cells were counted using a phase-contrast microscope. Five randomly selected high powered fields were counted for each membrane.
7
Transwell Invasion Assay for Cell Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transwell invasion assays were performed using a 24-well
polycarbonate membrane (Corning) with 8.0 μm pore size. The upper
chambers were precoated with 1 mg/ml Matrigel and incubated at 37 °C
for 2 h. The cells (5 × 104/well) in DMEM containing 1%
FBS were seeded into the upper chambers, and 600 μl of DMEM with 10%
FBS was added to the lower chambers. After incubation for 24 h at 37
°C, polycarbonate membranes were stained with HEMA3 staining kit
(Fisher). BT549 or 4T1 cells on the upper surface were removed with
cotton-tipped swabs. The number of invasive cells were counted from six
randomly selected visual fields using compound light microscopy (200x
magnification).
polycarbonate membrane (Corning) with 8.0 μm pore size. The upper
chambers were precoated with 1 mg/ml Matrigel and incubated at 37 °C
for 2 h. The cells (5 × 104/well) in DMEM containing 1%
FBS were seeded into the upper chambers, and 600 μl of DMEM with 10%
FBS was added to the lower chambers. After incubation for 24 h at 37
°C, polycarbonate membranes were stained with HEMA3 staining kit
(Fisher). BT549 or 4T1 cells on the upper surface were removed with
cotton-tipped swabs. The number of invasive cells were counted from six
randomly selected visual fields using compound light microscopy (200x
magnification).
8
Matrigel-Based Cell Invasion Assay
9
Transwell Invasion Assay for Cell Migration
10
Cytospins for Hematopoietic Cell Morphology
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!