Sections (4 µm) of formalin-fixed and paraffin-embedded specimens were deparaffinated followed by antigen retrieval using Target Retrieval (Dako, Glostrup, Denmark). The sections were subsequently blocked and stained in PBS plus 1% bovine serum albumin (Gibco, Carlsbad, CA, USA), 5% nonimmune donkey serum (Jackson ImmunoResearch, West Grove, PA, USA) and 0.1% Triton X-100. The following primary antibodies were used for immunofluorescence: anti-MRTFA (1:100; Sigma-Aldrich), anti-ACTA2-FITC (1:250; Sigma-Aldrich), anti-EPCAM (1:500; Cell Signaling Technology, Danvers, MA, USA), anti-CD45 (1:400; Cell Signaling Technology) and anti-CD31 (1:1000; Cell Signaling Technology). Human sections were imaged with a 20× Plan Apo DIC M objective (NA: 0.75) on a Nikon Ti-Eclipse (Nikon, Tokyo, Japan) inverted microscope equipped with an Andor Neo scMOS camera (Oxford Instruments, Abingdon, UK), a linear-encoded automated stage (Applied Scientific Instrumentation, Eugene, OR, USA) and a SOLA LED light engine (Lumencor, Beaverton, OR, USA) all run by NIS Elements software (Nikon).
Sola led light engine
Manufactured by Lumencor
Sourced in United States
The SOLA LED light engine is a compact, high-performance illumination source designed for use in laboratory and microscopy applications. It provides a stable, broad-spectrum white light output from a solid-state LED light source.
Automatically generated - may contain errors
Lab products found in correlation
Plan apo dic m objective, by Nikon (2 mentions)
Target retrieval, by Agilent Technologies (2 mentions)
Nis elements software, by Nikon (2 mentions)
Andor neo scmos camera, by Oxford Instruments (2 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Anti cd45, by Cell Signaling Technology (1 mentions)
Anti epcam, by Cell Signaling Technology (1 mentions)
Eclipse ti, by Nikon (1 mentions)
Anti cd31, by Cell Signaling Technology (1 mentions)
Eclipse ti e inverted microscope, by Nikon (1 mentions)
Orca flash4.0 digital cmos camera, by Hamamatsu Photonics (1 mentions)
Csu w1 confocal unit, by Yokogawa (1 mentions)
3 protocols using sola led light engine
1
Immunofluorescence Staining of FFPE Tissues
2
Immunohistochemical Detection of BMP1
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4 μm sections of formalin-fixed and paraffin-embedded specimens were deparaffinated followed by antigen retrieval using Target Retrieval (Dako #S1700) and quenching endogenous peroxidase activity. The sections were subsequently blocked with Vector Avidin Biotin Blocking Kit, stained in PBS plus 10% rabbit serum and 3% BSA with anti-BMP1 (0.5 µg/ml, R&D, #AF1297) for lung sections and secondary biotinylated antibodies, incubated with Vectastain ABC Elite reagent and Pierce Metal Enhanced DAB solution and counterstained with Mayer's hematoxylin. Human sections were imaged with a 20 × Plan Apo DIC M objective (NA: 0.75, Nikon) on a Nikon Ti-Eclipse inverted microscope equipped with an Andor Neo scMOS camera (Andor, Oxford Instruments), a linear-encoded automated stage (Applied Scientific Instrumentation), and a SOLA LED light engine (Lumencor) all run by NIS Elements software (Nikon).
3
Spinning-Disc Confocal Microscopy Protocol
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Imaging was performed using spinning-disc confocal microscopy with a 60 3 1.40 numerical aperture objective lens (Plan Apo l, Nikon, Tokyo, Japan). A CSU-W1 confocal unit (Yokogawa Electric Corporation, Tokyo, Japan) with five lasers (405, 488, 561, 640, and 685 nm, Coherent, Santa Clara, CA) and an ORCA-Flash 4.0 digital CMOS camera (Hamamatsu Photonics, Hamamatsu City, Japan) were attached to an ECLIPSE Ti-E inverted microscope (Nikon) with a perfect focus system. DNA images in Figures S2D andS2E or S3A were obtained using a SOLA LED light engine (Lumencor, Beaverton, OR).
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