Enucleated eye sections were stained with Hematoxylin, dehydrated, cleared, and permanently mounted for image acquisition on Nikon ECLIPSE 50i microscope. Immunofluorescence was performed as previously described31 (link). Snapshots of histology were taken using a Nikon ECLIPSE 50i microscope equipped with a 20x (numerical aperture 0.4) objective. Images were generated using an attached Rolera Bolt CMOS camera and QCapture Pro 7 software.
Eclipse 50i microscope
Manufactured by Nikon
Sourced in Japan, United States, Germany, France, Spain, China, United Kingdom
The Eclipse 50i microscope is an advanced optical instrument designed for laboratory use. It features high-quality optics and a robust construction to provide clear and detailed images of specimens. The microscope is capable of various observation techniques, including brightfield, darkfield, and phase contrast. Its core function is to enable detailed examination and analysis of samples across a range of scientific disciplines.
Automatically generated - may contain errors
Lab products found in correlation
Ds fi1 camera, by Nikon (9 mentions)
Ds ri1 camera, by Nikon (4 mentions)
Nis elements d 3, by Nikon (3 mentions)
Envision system hrp kit, by Agilent Technologies (2 mentions)
Fluoroshield mounting medium, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Ds 5mc, by Nikon (2 mentions)
Digital camera system ds fi1, by Nikon (2 mentions)
Nis elements f4, by Nikon (2 mentions)
Digital sight d5 u3, by Nikon (2 mentions)
Hematoxylin, by Thermo Fisher Scientific (2 mentions)
Streptavidin hrp, by Abcam (2 mentions)
Dab substrate kit, by Thermo Fisher Scientific (2 mentions)
Anti ki67, by Abcam (2 mentions)
Digital sight ds fi1 camera, by Nikon (2 mentions)
Dxm1200c digital camera, by Nikon (2 mentions)
Nis elements software, by Nikon (2 mentions)
Phorbol myristate acetate (pma), by Merck Group (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Axiovert microscope, by Zeiss (2 mentions)
228 protocols using eclipse 50i microscope
1
Histological Evaluation of Enucleated Eyes
2
Myelination and Histology Analysis of Spinal Cord
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Luxol fast blue staining was used to visualize myelin. Transverse SC cryostat sections were dehydrated by use of 85%, 95%, and 100% ethanol followed by xylene. They were incubated in 0.1% Luxol fast blue overnight at 56 °C (Thermo Fisher Scientific Inc.). The next day, the sections were differentiated in 0.05% lithium carbonate (Sigma Aldrich) and 70% ethanol, dehydrated with increasing concentrations of ethanol, and coverslipped using Permount (Thermo Fisher Scientific Inc.) mounting media. Pictures were obtained using a Nikon Eclipse 50i microscope equipped with a × 4 objective, using Ocular software (Digital Optics Ltd., Auckland, New Zealand).
Hematoxylin and eosin staining was performed with an automated stainer (Leica Autostainer XL, Nussloch, Germany). The SC sections were immersed in Harris hematoxylin and eosin stain. Following dehydration in 95% and 100% alcohol solutions, the section were coverslipped using Permount mounting media (Thermo Fisher Scientific Inc.) and pictures were taken with a Nikon Eclipse 50i microscope equipped with a × 10 objective and using the Ocular software (Digital Optics Ltd).
Hematoxylin and eosin staining was performed with an automated stainer (Leica Autostainer XL, Nussloch, Germany). The SC sections were immersed in Harris hematoxylin and eosin stain. Following dehydration in 95% and 100% alcohol solutions, the section were coverslipped using Permount mounting media (Thermo Fisher Scientific Inc.) and pictures were taken with a Nikon Eclipse 50i microscope equipped with a × 10 objective and using the Ocular software (Digital Optics Ltd).
3
Assessing Mitochondrial Network and Cell Death in C. elegans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the mitochondrial network, the CB5600 [ccIs4251 (Pmyo-3::Ngfp-lacZ; Pmyo-3::Mtgfp) I; him-8(e1489) IV] strain and CC91 [dys-1(eg33) I; ccIs4251 I; him-8(e1489) IV] strain were used for wt imaging and dystrophy imaging respectively, that uses myo-3p to express mitochondrial and nuclear GFP within all body-wall muscle cells. Worms were imaged at ×40 magnification using a Nikon Eclipse 50i microscope with images taken from the head and tail regions of each animal. Approximately 20 worms were imaged per strain, having been grown to Day 1 of adulthood either with or without SAA exposure. Animals were then classified into three mitochondrial network categories: well-networked, moderately networked and disorganised networks by visual analysis of mitochondrial morphology within each observable muscle cell, formulating individual percentages for each mitochondrial category, per worm. The same strains were used to evaluate cell death as described previously13 (link). Briefly, images were taken on Day 4 and Day 8 of adulthood using a Nikon Eclipse 50i microscope at ×10 magnification. After, manual counting of muscle cells for absent/aggregated GFP signals was performed per animal. C. elegans body-wall muscle cells are mononucleate, therefore, loss of observable nuclei is indicative of cell death as previously shown24 (link). Approximately 20 animals were assessed per condition.
4
Mitochondrial Imaging and Cell Death Analysis in C. elegans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial imaging was used in day 1 adults, with or without treatment, to examine the mitochondrial network. The CB5600 [ccIs4251 (Pmyo-3::Ngfp-lacZ; Pmyo-3::Mtgfp) I; him-8(e1489) IV] strain and CC91 [dys-1(eg33) I; ccIs4251 I; him-8(e1489) IV] were used for WT imaging and dystrophy imaging, respectively. Worms were cultured on NaGYY as described. Approximately 20 day 1 adults were picked into 20 µL of M9 buffer on a microscope slide with a coverslip applied. Worms were imaged at 40× magnification using a Nikon Eclipse 50i microscope. CB5600 and CC91 animals were also used for the cell-death images. The protocol used was as described (20 (link)). Briefly animals were assessed at day 4 and day 8 of adulthood, where the number of dead muscle cells was determined by quantifying the number of muscle cells that had lost their distinct circular nuclear GFP signal. Approximately 30 animals were picked into 20 µL of M9 buffer on a microscope slide with a coverslip applied. Worms were imaged at 10x magnification by using a Nikon Eclipse 50i microscope.
5
Immunofluorescent Localization of Plin1 and Plin2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed for 20 min with 4% paraformaldehyde and 0.01% Triton X-100 in PBS, followed by 3 rinses with PBS for 5 min each. Nonspecific binding sites in cells were blocked with 5% donkey serum for 60 min. The cells were incubated with polyclonal antibodies against Plin1 or Plin2 [32 (link)] at 1:500 overnight at 4°C, then with FITC-conjugated secondary antibody at 1:500 for 1 h. Cell nuclei were stained with Hoechst 33258. Immunofluorescent signals were observed under a Nikon Eclipse 50i microscope.
6
Sperm Membrane Integrity Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The specimens were stained using the Live/Dead Sperm Viability Kit (Molecular Probes Inc., Leiden, The Netherlands). A 5 μL volume of 50× diluted SYBR-14 was added to 1 mL of diluted ejaculate, followed by incubation at 36 °C for 10 min. Then 5 μL of propidium iodide (PI) was added, followed by incubation at 36 °C for 10 min. A drop of solution was applied to a heated microscope slide, and the integrity of the cell membranes was examined using a Nikon Eclipse 50i microscope with a fluorescence. One slide per sample were analyzed. On each slide, 200 sperm were evaluated. Sperm emitting green fluorescence over the entire head were identified as live cells (with an intact cell membrane stained by SYBR-14), sperm emitting red fluorescence over the entire head or on part of the head and sperm emitting yellow-orange fluorescence over the entire head were identified as dead (with a damaged cell membrane, stained by PI), and sperm emitting yellow-orange fluorescence over the entire head were identified as moribund sperm.
7
Teratoma Formation and Lineage Differentiation of iPSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For in vivo differentiation of iPS Cells: 1×106 cells were resuspended in 50 μL 1:3 diluted GFR matrigel (BD Biosciences). The cell suspension was then injected into the left testis of a severe combined immunodeficiency (SCID) mouse, while the right testis was injected with 1:3 diluted GFR matrigel alone, as a negative control. Following teratoma formation, classic histological staining was carried out using Mayer’s Haematoxylin and Eosin. Images were taken using a Zeiss Axioscope Z Plus microscope.
For in vitro differentiation of iPS cells: iPS cells were grown for 10 days in DMEM supplemented with 20% FBS. Medium was changed once every three days, and cells were then fixed with 4% PFA. The following antibodies were used: anti-α-fetoprotein (endoderm), anti-βIII-tubulin (ectoderm), and anti-α-smooth muscle actin (mesoderm), all from Millipore. Images were taken using a Nikon Eclipse 50i microscope at x10 magnification.
For in vitro differentiation of iPS cells: iPS cells were grown for 10 days in DMEM supplemented with 20% FBS. Medium was changed once every three days, and cells were then fixed with 4% PFA. The following antibodies were used: anti-α-fetoprotein (endoderm), anti-βIII-tubulin (ectoderm), and anti-α-smooth muscle actin (mesoderm), all from Millipore. Images were taken using a Nikon Eclipse 50i microscope at x10 magnification.
8
Macrophage Foam Cell Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
THP-1 cells were grown in media and treated with 10 ng/mL phorbol myristate acetate (MilliporeSigma, Temecula, CA, USA) to induce differentiation into macrophage for 48 h. The culture medium was subsequently changed to RPMI and 1% lipid-depleted fetal bovine serum (FBS) from differential ultracentrifugation, as described previously [51 (link)]. Then, the macrophages were treated 50 μg/mL LDL with 50 μg/mL H1 or H5 at 37 °C for 48 h. Foam cell formation in macrophages was examined by using Oil red O staining [52 (link)]. The images for foam cell formation were observed in an ECLIPSE 50i microscope (Nikon, Tokyo, Japan) and ECHO Retrieval system (ECHO, San Diego, CA, USA). The percentage of foam cell formation was quantitated by dividing the number of Oil red O staining macrophages by the total number of macrophages in 3 random microscopic fields (over 200 cells).
9
Fluorescence Imaging of Probes in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All fluorescence microscopy images and corresponding brightfield images were obtained on an Olympus IX73 microscope. All probe 2 cell images were obtained using a filter cube consisting of 380–420 nm excitation bandpass filter, a 484 nm dichroic mirror, and a 513–604 nm emission bandpass filter. Cell images of 3 were acquired using a filter cube comprising 503–557 nm excitation bandpass filter, a 580 nm dichroic mirror, and a 600–700 nm emission bandpass filter. All cells were plated in 8-chambered wells (Lab-Tek® Chambered #1.0 Borosilicate Coverglass System) at a density of 5 × 104 cells per well one day before imaging. The cells were washed once with DPBS, and were incubated in Opti-MEM with or without 100 μM LPA for 1 h. 20 μM probe 2 was then diluted into the appropriate wells and the cells were incubated for 3 h. The cells were washed five times in DPBS and were imaged in DPBS. All images were analysed using Olympus CellSens Dimension V1.18. H&E images were acquired from a Nikon Eclipse 50i Microscope with a Nikon DS-Ri1 camera and NIS-Elements BR3.0 software.
10
Quantifying MN-miRNA Uptake in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MN-miRNA uptake by the human cancer cell lines MDA-MB-231 and T98G was assessed through fluorescence imaging. T98G cells (ATCC) were seeded at a density of 1 × 105 cells per well in a 12-well glass plate. MN-miR4261 was added into each well at various concentrations ranging from 0, 5 and 30 μM and incubated at 37 °C in a humidified 6% CO2 atmosphere for 48 hrs. The cells were fixed in 4% paraformaldehyde and mounted on slides with Vectashield Mounting Media with DAPI for nuclear staining. Microscopic images of Cy5.5 and DAPI were obtained using a Nikon Eclipse 50i microscope, and then raw images were imported by ImageJ and processed to generate an overlayed image, showing the subcellular location of the MN-MDRmiR. Following the same procedure, MDA-MB-231 was incubated with MN-miR4539 and the cellular uptake and localization of MN-MDRmiR in the cytosol was visualized following the same procedures described above.
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!