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Axioimager m1

Manufactured by Hamamatsu Photonics
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The AxioImager M1 is a light microscope designed for biological and material science applications. It features a compact and ergonomic design, and supports a range of imaging techniques including bright-field, phase contrast, and differential interference contrast (DIC) microscopy.

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Lab products found in correlation

Anti rabbit igg alexafluor 594, by Thermo Fisher Scientific (2 mentions) Volocity 4, by PerkinElmer (2 mentions) Mouse anti cyclin a, by Santa Cruz Biotechnology (2 mentions) Mouse anti α tubulin, by Merck Group (2 mentions) Alexa fluor 555, by Thermo Fisher Scientific (2 mentions) Mouse anti histone h2ax pser139, by Merck Group (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Mouse anti flag, by Merck Group (2 mentions) Rabbit anti gfp, by Abcam (1 mentions) Duolink in situ detection reagents red, by Olink (1 mentions) Volocity software, by PerkinElmer (1 mentions)

6 protocols using axioimager m1

1

Immunofluorescent Detection of 53BP1

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Cells were washed with PBS and fixed with 4% formaldehyde for 10 min at room temperature, permeabilized with 0.3% Triton X-100 in PBS for 5 min at room temperature and then blocked with 3% BSA/10% FBS in PBS for 1 h at room temperature. Samples were then incubated with rabbit anti-53BP1 overnight at 4°C, washed with 0.05% PBS-Tween20 and incubated with anti-rabbit IgG AlexaFluor 594 (Molecular Probes). DNA was counterstained with DAPI and images were acquired using a Zeiss AxioImager M1, using a Hamamatsu digital camera and the Volocity 4.3.2 software (Perkin Elmer).
Sarek G., Kotsantis P., Ruis P., Van Ly D., Margalef P., Borel V., Zheng X.F., Flynn H.R., Snijders A.P., Chowdhury D., Cesare A.J, & Boulton S.J. (2019). CDK phosphorylation of TRF2 controls t-loop dynamics during the cell cycle. Nature, 575(7783), 523-527.
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2

Immunofluorescent Detection of 53BP1

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Cells were washed with PBS and fixed with 4% formaldehyde for 10 min at room temperature, permeabilized with 0.3% Triton X-100 in PBS for 5 min at room temperature and then blocked with 3% BSA/10% FBS in PBS for 1 h at room temperature. Samples were then incubated with rabbit anti-53BP1 overnight at 4°C, washed with 0.05% PBS-Tween20 and incubated with anti-rabbit IgG AlexaFluor 594 (Molecular Probes). DNA was counterstained with DAPI and images were acquired using a Zeiss AxioImager M1, using a Hamamatsu digital camera and the Volocity 4.3.2 software (Perkin Elmer).
Sarek G., Kotsantis P., Ruis P., Van Ly D., Margalef P., Borel V., Zheng X.F., Flynn H.R., Snijders A.P., Chowdhury D., Cesare A.J, & Boulton S.J. (2019). CDK phosphorylation of TRF2 controls t-loop dynamics during the cell cycle. Nature, 575(7783), 523-527.
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3

Immunofluorescence Staining of HeLa and Flp-In T-REx 293 Cells

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HeLa Kyoto cells or Flp-In T-REx 293 cells were fixed and processed for immunofluorescence as described45 (link). In brief, cells were fixed in 4% paraformaldehyde for 10 min at the room temperature followed by ice-cold methanol for 1 min. The cells were then wash with 1 × phosphate-buffered saline (PBS) and permeabilized with 0.5% Triton X-100 for 5 min before antibody staining. The primary antibodies used were as follows: mouse anti-histone H2AX pSer139 (1:1,000, Millipore 05-636-1), mouse anti-α-tubulin (1:5,000, Sigma-Aldrich T9026), rabbit anti-GFP (1:5,000, Abcam ab290), mouse anti-cyclin A (1:200, Santa Cruz sc-56299), mouse anti-FLAG (1:1,000, Sigma-Aldrich A8592). Secondary antibodies conjugated to Alexa Fluor 488 and Alexa Fluor 555 (1:1,000, Molecular Probes) were used for detection. DNA was stained with 4,6-diamidino-2-phenylindole. Images were acquired using Zeiss AXIO Imager M1 with a × 40 EC-Plan-Neofluor lens and Hamamatsu photonics camera under the control of Volocity software. Images were processed using Adobe Photoshop.
Chan Y.W, & West S.C. (2014). Spatial control of the GEN1 Holliday junction resolvase ensures genome stability. Nature Communications, 5, 4844.
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4

Immunofluorescence Imaging of Cell Markers

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HeLa Kyoto cells or Flp-In T-REx 293 cells were fixed and processed for immunofluorescence as described45 (link). In brief, cells were fixed in 4% paraformaldehyde for 10 min at the room temperature followed by ice-cold methanol for 1 min. The cells were then wash with 1xPBS and permeabilised with 0.5% Triton X-100 for 5 min before antibody staining. The primary antibodies used were: mouse anti-histone H2AX pSer139 (1:1000, Millipore 05-636-1), mouse anti-α-tubulin (1:5000, Sigma-Aldrich T9026), rabbit anti-GFP (1:5000, Abcam ab290), mouse anti-cyclin A (1:200, Santa Cruz sc-56299), mouse anti-FLAG (1:1000, Sigma-Aldrich A8592). Secondary antibodies conjugated to Alexa Fluor 488 and Alexa Fluor 555 (1:1000, Molecular Probes) were used for detection. DNA was stained with DAPI. Images were acquired using Zeiss AXIO Imager M1 with a 40x EC-Plan-Neofluor lens and Hamamatsu photonics camera under the control of Volocity software. Images were processed using Adobe Photoshop.
Chan Y.W, & West S.C. (2014). Spatial control of the GEN1 Holliday junction resolvase ensures genome stability. Nature Communications, 5, 4844.
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5

Proximity Ligation Assay for RNAPII-PCNA Interaction

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Cells grown on coverslips were pre-extracted in 0.5% NP40 on ice for 4min then washed once with PBS an fixed with 4% formaldehyde in PBS for 15 min at room temperature, washed three times with PBS, blocked with 3% BSA/10% fetal bovine serum and incubated with antibodies mouse RNAPII 8WG16Pol 1:200 and rabbit PCNA 1:200 overnight at 4°C. PLA was performed following the manufacturer’s instructions using the Duolink anti-Mouse MINUS and anti-Rabbit PLUS In Situ PLA probes and the Duolink In Situ Detection Reagents Red (Olink Bioscience). Images were acquired using a Zeiss AxioImager M1, equipped with a Hamamatsu digital camera and the Volocity software (Perkin Elmer). PLA foci were analyzed using ImageJ (https://imagej.nih.gov/ij/).
Kotsantis P., Segura-Bayona S., Margalef P., Marzec P., Ruis P., Hewitt G., Bellelli R., Patel H., Goldstone R., Poetsch A.R, & Boulton S.J. (2020). RTEL1 Regulates G4/R-Loops to Avert Replication-Transcription Collisions. Cell Reports, 33(12), 108546.
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6

Imaging Biofilms in Microtiter Plates

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About 1 μl aliquots of media from 96-well plates were placed on glass microscope slides and imaged using a Zeiss AxioImager M1 with a Hamamatsu Orca 03 12-bit grayscale digital color camera. Images were taken at 40× and 100×. Nile Red was detected using the Texas Red filter. To image intact biofilms within each well in glass-bottom 96-well microtiter plates, we used a Zeiss AxioObserver Z1 Live-Cell system with a QImaging Retiga SRV and a QImaging 5MPix Micropublisher camera. Images were taken with a 0.01–0.05 s exposure time (depending on magnification) and analyzed using the Zeiss Axio Vision software.
French K.E, & Terry N. (2019). A High-Throughput Fluorescence-Based Assay for Rapid Identification of Petroleum-Degrading Bacteria. Frontiers in Microbiology, 10, 1318.
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