For immunohistochemistry staining, heart cryosections were fixed with 4% paraformaldehyde, permeabilized and blocked with Protein Block Solution (DAKO, Carpinteria, CA) containing 0.1% saponin (Sigma, St Louis, MO), and then incubated with the following antibodies overnight at 4 °C: mouse anti-alpha sarcomeric actin (1:100, a7811, Sigma), rabbit anti-CD45 (1:100, ab10559, Abcam, Cambridge, United Kingdom), mouse anti-Actin, α-Smooth Muscle antibody (1:100, A5228, Sigma), rabbit anti-Ki67 (1:100, ab15580, Abcam), rabbit anti-CD3 (1:100, ab16669, Abcam) and mouse anti-CD8 alpha (1:100, mca48r, abd Serotec, Raleigh, NC ). FITC- or Texas-Red secondary antibodies (1:100) were obtained from Abcam Company and used for the conjunction with these primary antibodies. For assessment of cell apoptosis, heart cryosections were incubated with TUNEL solution (Roche Diagnostics GmbH, Mannheim, Germany) and counter-stained with DAPI (Life Technology, NY, USA). For assessment of angiogram, heart cryosections were incubated with Lectin (FL-1171, Vector laboratories, Burlingame, CA, USA). Images were taken by an Olympus epi-fluorescence microscopy system.
Epi fluorescence microscopy system
Manufactured by Olympus
The Epi-fluorescence microscopy system is a versatile imaging tool designed for visualizing fluorescently labeled samples. It utilizes epifluorescence illumination, where the excitation light is directed through the objective lens and onto the sample, allowing for efficient detection of fluorescent signals. This system is capable of capturing high-quality images of a wide range of fluorescent samples, making it a valuable tool for various applications in life science research and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Ab16669, by Abcam (4 mentions)
Protein block solution, by Agilent Technologies (4 mentions)
Mca48r, by Bio-Rad (4 mentions)
A7811, by Merck Group (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions)
Saponin, by Merck Group (4 mentions)
Ab15580, by Abcam (3 mentions)
A5228, by Merck Group (3 mentions)
Tunel solution, by Roche (3 mentions)
Ab10559, by Abcam (2 mentions)
Fitc or texas red secondary antibodies, by Abcam (2 mentions)
Ab955, by Abcam (2 mentions)
Ab6994, by Abcam (1 mentions)
Fitc conjugated secondary antibody, by Abcam (1 mentions)
5 protocols using epi fluorescence microscopy system
1
Immunohistochemical Analysis of Cardiac Tissue
2
Immunophenotyping and Apoptosis Analysis of Cardiac Tissue
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Heart cryosections were
fixed with 4% paraformaldehyde, permeabilized, and blocked with protein
block solution (DAKO, Carpinteria, CA) with 0.1% saponin (Sigma) and
then incubated with the primary antibodies overnight at 4 °C.
Primary antibodies were listed as follows: rabbit anti-CD3 (ab16669,
Abcam, Cambridge, United Kingdom), mouse anti-CD8 alpha (mca48r, abd
Serotec, Raleigh, NC), mouse anti-CD68 (ab955, Abcam), mouse anti-alpha
sarcomeric actin (a7811, Sigma), rabbit anti-Ki67 (ab15580, Abcam),
rabbit anti-vWF (ab6994, Abcam), and a smooth muscle actin antibody
(A5228, Sigma). FITC- or Texas-Red secondary antibodies obtained from
Abcam Company were incubated and conjoined with related primary antibodies.
For evaluation of cell apoptosis, heart cryosections were incubated
with TUNEL solution (Roche Diagnostics GmbH, Mannheim, Germany) and
counter-stained with DAPI (Life Technology, NY, USA). Images were
taken by an Olympus epi-fluorescence microscopy system as previously
described.48 (link),49 (link)
fixed with 4% paraformaldehyde, permeabilized, and blocked with protein
block solution (DAKO, Carpinteria, CA) with 0.1% saponin (Sigma) and
then incubated with the primary antibodies overnight at 4 °C.
Primary antibodies were listed as follows: rabbit anti-CD3 (ab16669,
Abcam, Cambridge, United Kingdom), mouse anti-CD8 alpha (mca48r, abd
Serotec, Raleigh, NC), mouse anti-CD68 (ab955, Abcam), mouse anti-alpha
sarcomeric actin (a7811, Sigma), rabbit anti-Ki67 (ab15580, Abcam),
rabbit anti-vWF (ab6994, Abcam), and a smooth muscle actin antibody
(A5228, Sigma). FITC- or Texas-Red secondary antibodies obtained from
Abcam Company were incubated and conjoined with related primary antibodies.
For evaluation of cell apoptosis, heart cryosections were incubated
with TUNEL solution (Roche Diagnostics GmbH, Mannheim, Germany) and
counter-stained with DAPI (Life Technology, NY, USA). Images were
taken by an Olympus epi-fluorescence microscopy system as previously
described.48 (link),49 (link)
3
Immunostaining of Cardiac Cryosections
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Heart cryo‐sections were fixed with 4% paraformaldehyde, permeabilized and blocked with protein block solution (DAKO, Carpinteria, CA, USA) containing 0.1% saponin (Sigma‐Aldrich) and then incubated with the following antibodies overnight at 4°C: mouse anti‐alpha sarcomeric actin (1:100, a7811, Sigma‐Aldrich) and human nuclei antigen (HNA) (1:200, Millipore, Billerica, MA), followed by incubation with Texas‐Red or FITC‐conjugated secondary antibodies. The images were taken by an Olympus epi‐fluorescence microscopy system.
4
Immunohistochemical Analysis of Heart Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Example 11
Histology:
For immunohistochemistry staining, heart cryosections were fixed with 4% paraformaldehyde, permeabilized and blocked with Protein Block Solution (DAKO, Carpinteria, Calif.) containing 0.1% saponin (Sigma, St. Louis, Mo.), and then incubated with the following antibodies overnight at 4° C.: mouse anti-alpha sarcomeric actin (1:100, a7811, Sigma), rabbit anti-CD45 (1:100, ab10559, Abcam, Cambridge, United Kingdom), mouse anti-Actin, α-Smooth Muscle antibody (1:100, A5228, Sigma), rabbit anti-Ki67 (1:100, ab15580, Abcam), rabbit anti-CD3 (1:100, ab16669, Abcam) and mouse anti-CD8 alpha (1:100, mca48r, abd Serotec, Raleigh, N.C.). FITC- or Texas-Red secondary antibodies (1:100) were obtained from Abcam Company and used for the conjunction with these primary antibodies. For assessment of cell apoptosis, heart cryosections were incubated with TUNEL solution (Roche Diagnostics GmbH, Mannheim, Germany) and counter-stained with DAPI (Life Technology, NY, USA). For assessment of angiogram, heart cryosections were incubated with Lectin (FL-1171, Vector laboratories, Burlingame, Calif., USA). Images were taken by an Olympus epi-fluorescence microscopy system.
5
Immune Cell Infiltration After Cardiac Infarction
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