Images were acquired with Eclipse E800 (Nikon). Additional images were captured with a Leica DM LB compound microscope (Leica Microsystems). All the pictures were focused on the first lumbar intervertebral disc. For fluorescent images, frozen sections were dehydrated at 4°C overnight and wash in 1X PBS then overlayed with coverslips onto Vectashield Hard Set mounting medium (Vector Laboratories, Burlingame, CA, USA). Photos were taken using filters for Red Fluorescent Protein (RFP) and fluorescein isothiocyanate (FITC).
Dm lb compound microscope
Manufactured by Leica
Sourced in United States
The Leica DM LB compound microscope is a versatile and high-quality optical instrument designed for a wide range of laboratory applications. It features a sturdy construction, advanced optics, and user-friendly controls to provide a reliable and efficient tool for microscopic observations.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse e800, by Nikon (2 mentions)
Vectashield hardset mounting medium, by Vector Laboratories (2 mentions)
Anti gfp staining, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
Microm hm 355, by Thermo Fisher Scientific (1 mentions)
M165 fc, by Leica (1 mentions)
Application suite, by Leica (1 mentions)
Cv1000 spinning disk confocal microscope, by Olympus (1 mentions)
Vectorshield, by Vector Laboratories (1 mentions)
Smz1500, by Nikon (1 mentions)
S6d dissecting microscope, by Leica (1 mentions)
Pgem t easy, by Promega (1 mentions)
11 protocols using dm lb compound microscope
1
Lumbar Disc Fluorescence Imaging
2
Histomorphometric Analysis of Bone Tumor
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Hind limbs were fixed for 48 hours in 10% neutral buffered formalin, decalcified in 10% ethylenediaminetetraacetic acid for 2 weeks, processed and embedded in paraffin wax. An automated microtome (HM 355S; MICROM International, Walldorf, Germany) was used to cut 3.5-μm-thick, longitudinal, midsagittal sections of the embedded tissue. Sections were stained with hematoxylin and eosin (H&E) and TRAP (for the identification of osteoclasts) for histomorphometric analysis. Images were captured with a Leica DM LB compound microscope (Leica Microsystems, Buffalo Grove, IL, USA) and a QImaging MicroPublisher cooled charge-coupled device color digital camera (QImaging, Surrey, BC, Canada). MetaMorph software was used to analyze total tumor burden and osteoclast number in multiple sections from the femora and tibiae of the hind limbs. Tumor burden was defined as the area of bone occupied by the tumor at 50× magnification of H&E-stained sections [27 (link)]. Osteoclast number per millimeter of bone surface at the interface with tumor was measured from TRAP-stained tibiae at 200× magnification.
3
Fluorescent Microscopy Protocol for Tissue Imaging
4
Histomorphometric Analysis of Bone Tumor
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Bones from the forelimbs and hindlimb were collected upon euthanasia, fixed in 10% formalin for 48h, decalcified in 10% EDTA for 2 weeks, and embedded in paraffin. Tissue sections from tibia and femur were cut using an automated Microm HM 355S microtome (Thermo Fisher Scientific) and stained with hematoxylin and eosin (H&E) or for TRAP (tartrate-resistant acid phosphatase) activity. Bright field images were captured using a Q-Imaging Micropublisher Cooled CCD color digital camera (Vashaw Scientific Inc., Washington, DC, USA) on a Leica DM LB compound microscope (Leica Microsystem, Bannockburn, IL, USA). Histomorphometric analysis of tumor burden area, defined as area of bone occupied by cancer cells, and total bone area was performed using BIOQUANT OSTEO software (Image Analysis Corporation). Osteoclast numbers at the tumor bone interface were quantified on TRAP-stained sections of femur and tibia at 400X magnification.
5
Binocular Stereomicroscopy for ISH
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A Leica M16 2F binocular stereomicroscope was used to observe the embryos labeled by in situ hybridizations. For cryo-sections ISH, the sections were observed under a Leica DMLB compound microscope. All images were taken by a camera color CoolSnapPro and treated using Adobe Photoshop Elements 9 (Adobe, CA, USA) for contrast and brightness.
6
Dissection and Imaging of Drosophila Tissues
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Adult flies were partially dissected then mounted directly onto plasticine and monitored with a Leica MZ6 stereomicroscope. All eye images were recorded on a Leica DC300n digital camera and Leica Application Suite.
Midguts, salivary glands and the brain were dissected from third instar larvae in cold phosphate buffered saline (PBS) and mounted directly onto glass slides in Vectorshield (Vectorlabs) or fixed in 4% paraformaldehyde in PBS. Monoclonal α-Prospero and α–Fasciclin I primary antibodies used for Zip88E>mCD8::eGFP co-localization studies were obtained from the Developmental Studies Hybridoma Bank and used at 1:50 and 1:200 dilution respectively, followed by α-mouse AlexaFluor568 secondary antibody (Molecular Probes) used at 1:1000 dilution. MtnB:EYFP fluorescence in larval tissue was viewed on a Leica M165 FC dissecting microscope using a Leica DFC450 camera and Leica Application Suite. Higher magnification imaging was performed on either: 1) a Leica DMLB compound microscope using a Leica DC300 camera and Leica Application Suite at a magnification of 10x and 20x; or 2) an Olympus CV1000 spinning disk confocal microscope with a 10x dry objective lens or a 60x immersion objective lens.
Midguts, salivary glands and the brain were dissected from third instar larvae in cold phosphate buffered saline (PBS) and mounted directly onto glass slides in Vectorshield (Vectorlabs) or fixed in 4% paraformaldehyde in PBS. Monoclonal α-Prospero and α–Fasciclin I primary antibodies used for Zip88E>mCD8::eGFP co-localization studies were obtained from the Developmental Studies Hybridoma Bank and used at 1:50 and 1:200 dilution respectively, followed by α-mouse AlexaFluor568 secondary antibody (Molecular Probes) used at 1:1000 dilution. MtnB:EYFP fluorescence in larval tissue was viewed on a Leica M165 FC dissecting microscope using a Leica DFC450 camera and Leica Application Suite. Higher magnification imaging was performed on either: 1) a Leica DMLB compound microscope using a Leica DC300 camera and Leica Application Suite at a magnification of 10x and 20x; or 2) an Olympus CV1000 spinning disk confocal microscope with a 10x dry objective lens or a 60x immersion objective lens.
7
Histological Analysis of Bone Tumor Localization
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Hind limbs were fixed as described above and decalcified in 10% EDTA for two weeks. Tissues were processed as previously described (18 (link)) and sections were stained with hematoxylin and eosin, and tartrate resistant alkaline phosphatase (TRAP). Tumor localization was confirmed with anti-GFP staining (Abcam). Sections were viewed on a Leica DM LB compound microscope outfitted with a Q-Imaging Micropublisher Cooled CCD color digital camera (W. Nuhsbaum Inc, McHenry, IL).
8
Histological Analysis of Bone Tumor Localization
9
Histological Analysis of Decalcified Hind Limbs
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The hind limbs were fixed with formalin and decalcified in 10% EDTA for 2 weeks. The tissues were processed as previously described [22 (link)], and the sections stained with hematoxylin and eosin, and tartrate-resistant alkaline phosphatase (TRAP) (Sigma-Aldrich, St. Louis, MO, USA). The sections were scored on a Leica DM LB compound microscope outfitted with a Q-Imaging Micropublisher Cooled CCD color digital camera (W. Nuhsbaum Inc., McHenry, IL, USA).
10
Larval Morphology and Habitat Analysis of Stenhelmoides rufulus
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Specimens were captured with a Surber net, killed and preserved in a 75% ethyl alcohol solution. Larval specimens were cleared using warm lactic acid and dissected and mounted on glass slides with PVLG medium. Observations were made using a Leica S6D dissecting microscope and a Leica DMLB compound microscope, both equipped with photographic cameras. Drawings of setae were scanned and digitally edited. Additional photographs (Fig. 1D,F,G ) were taken with a digital camera Nikon DS-Ri1 U3, adapted to a stereoscope Nikon SMZ-1500. The photos were assembled using the freeware programs CombineZP24 and Helicon Focus®. The larval morphology nomenclature of Harris25 , Shepard26 , Borror et al.27 , Lawrence28 and Kodada et al.1 were followed for the descriptions. Data on physical and chemical parameters of Stenhelmoides habitats were provided by the Water Laboratory of Institute for Research and Development in Water Supply, Environmental Sanitation and Water Resource Conservation - CINARA (by its acronym in Spanish) of Universidad del Valle (CINARA, unpublished data).![]()
Habitus and color variation of Stenhelmoides rufulus larvae. (
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