The largest database of trusted experimental protocols

Lionheart microscope

Manufactured by Agilent Technologies
Sourced in United States

The Lionheart microscope is a compact, automated microscope designed for routine imaging and analysis of cells and small samples. It features a motorized stage, autofocus, and LED illumination to capture high-quality images. The Lionheart microscope is suitable for a variety of applications, including cell culture monitoring, fixed-cell imaging, and time-lapse imaging.

Automatically generated - may contain errors

11 protocols using lionheart microscope

1

Immunofluorescence Staining of pMLKL and IL-8

Check if the same lab product or an alternative is used in the 5 most similar protocols
Frozen sections (5 μm) were fixed with 4% paraformaldehyde-PBS 20 min at room temperature, permeabilized with 0.3% Triton X-100 in 1X PBS, blocked in 3% BSA (or 10% FBS) in 1X PBS, stained with primary antibodies for overnight at 4°C, followed by staining with 2° antibodies (1 hr) and DAPI stain. Samples were covered with coverglasses and imaged using Lionheart microscope (Biotek). S358 pMLKL monoclonal antibody (Abcam, EPR9514, 1:500) and human IL-8 (CXCL8) Monoclonal Antibody (3IL8-H10, Thermo Fisher, 1:1000) were used to perform the immunofluorescence staining. Samples were imaged using Lionheart microscope (Biotek).
+ Open protocol
+ Expand
2

Cell-Material Direct Interaction Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
By performing this assay, the cells were prepared as in the cell-attachment method. For this assay, the cells were seeded into 24-well plates alongside previously placed test materials. In this method, the direct interaction between the cells and the test material was checked by evaluating the morphology and staining as in the previous method, but only the percentage of live and dead cells was evaluated on a BioTek Lionheart microscope. To ensure that no direct cytotoxic effects were observed, the samples in the co-culture assay were incubated for 72 h.
+ Open protocol
+ Expand
3

Culturing and Validating Cell Lines for Hypoxia Studies

Check if the same lab product or an alternative is used in the 5 most similar protocols
All cell lines, with the exception of SUMs, were obtained from the ATCC. SUMs were purchased from Asterand Bioscience. Cells were cultured per company provided protocols. The MCF10A and MCF10A ER-expressing cells were a kind gift from Ben Ho Park and cultured as previously described (27 (link)). CRISPR edited MCF-7 HIF-1α, HIF-2α and control knockout cell lines were previously generated in our laboratory (8 (link)). All cell lines used in the study were authenticated by STR sequencing and confirmed to be mycoplasma free. Cells were maintained in a humidified environment at 37°C and 5% CO2 during culture and live-cell imaging. Hypoxic cells were maintained at 37°C in an invivo 200 hypoxia workstation equipped with a digitally controlled oxygen regulator and maintained at 1% O2, 5% CO2, and 94% N2. Live-cell microscopy experiments were conducted in a McCoy incubator maintained at 1% O2, 5% CO2, and 94% N2 and imaged with a Lionheart microscope (Biotek).
+ Open protocol
+ Expand
4

Airway Inflammation and Mucus Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
Left lung lobes were inflated with 10% neutral buffered formalin at 25 cm H2O and paraffin embedded. Slides were stained with H&E to assess immune cell infiltration/aggregation in peribronchial and perivascular spaces (0–4 scale) and inducible bronchial-associated lymphoid tissue (iBALT) formation. Slides were stained with Alcian Blue-Periodic Acid Schiff (PAS) to quantify mucous cells in the airway epithelium. Photomicrographs of four different airways were taken at 100X on Lionheart microscope (Biotek, Winooski, VT). PAS positive and negative cells were quantified blinded using ImageJ (NIH) and reported as a percentage of total airway epithelial cells counted.
+ Open protocol
+ Expand
5

Immunofluorescence Staining of Activated Caspase-3

Check if the same lab product or an alternative is used in the 5 most similar protocols
Frozen sections (5 μm) were fixed with 4% paraformaldehyde-PBS 20 min at room temperature, permeabilized with 0.3% Triton X-100 in 1X PBS, blocked in 3% BSA (or 10% FBS) in 1X PBS, stained with primary antibodies for overnight at 4°C, followed by staining with 2° antibodies (1 hr) and DAPI stain. Samples were covered with coverglasses and imaged using Lionheart microscope (Biotek). Ac-caspase-3 monoclonal antibody (9661S, Cell Signaling, 1:200) was used to perform the immunofluorescence staining.
+ Open protocol
+ Expand
6

Quantifying Protein Synthesis in C. albicans

Check if the same lab product or an alternative is used in the 5 most similar protocols
To observe translation in C. albicans cells, we used a Click-iT Protein Synthesis Assay Kit (Thermofisher) per the manufacturer’s instructions. WT C. albicans was subcultured from overnight cultures in YPD into minimal media and allowed to grow to log phase (OD600 0.4 – 0.8). After this growth, non-control cultures were treated with 25 μM NSC 697923 for 10 minutes. 50 μM HPG Reagent was used. Cells were fixed with 4% Paraformaldehyde (PFA) and permeabilized with 0.5% Triton X-100 in PBS. To quantify translation, cells were imaged on a Biotek Lionheart microscope at 40X magnification using a Texas Red channel at the same exposure time for each image. The same cells were also examined by flow cytometry on a BD Fortessa flow cytometer and analyzed using FloJo (BD Biosciences).
+ Open protocol
+ Expand
7

Immunostaining of Cells after Plasma Membrane Damage

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were plated on 2-well or 4-well glass chamber slides (Thermo Fisher) coated with fibronectin (100 μg/mL in PBS, Millipore). Cells were maintained in complete media at 37°C and 5% CO2 in an environmentally controlled chamber. After PM damage, cells were fixed cells in 4% paraformaldehyde-PBS 10~20 min at room temperature, permeabilized with 0.3% Triton X-100 in 1X PBS, blocked in 3% BSA in 1X PBS, stained with primary antibodies for 2–4 hr (room temperature) to overnight at 4°C, followed by staining with 2° antibodies (1 hr) and DAPI stain. Samples were imaged using Lionheart microscope (Biotek).
+ Open protocol
+ Expand
8

Murine Pancreatic Tumor Tissue Staining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Murine pancreatic tumor tissue slides were sectioned (5 microns) from the paraffin-embedded blocks. Tissues were then stained following standard procedures. Briefly, the slides were first deparaffinized with xylene and rehydrated using a gradient of ethanol and distilled water. An antigen retrieval step was performed in 10 mM sodium citrate buffer, at pH 6, in the presence of 0.05% Tween 20. The slides were treated with 3% H2O2 for 5 min prior to blocking for 20 min with horse serum at room temperature (RT). Antibody incubation was performed overnight (O/N) at 4 °C for the primary antibody and for 1 h at RT for the secondary antibody. After several washes with TBS-containing 0.05% Tween 20 (TBS-T), the slides were incubated with Vector SG substrate followed by nuclear fast red (Vector Laboratories; Burlingame, CA, USA). A coverslip with permanent mounting media was added to the slides prior to imaging. Images were taken on a Lionheart microscope (BioTek, Winooski, VT, USA).
+ Open protocol
+ Expand
9

Immunofluorescence Imaging of Cultured Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells grown on 1.5 mm thick glass bottom dishes were fixed in 4% paraformaldehyde for 10 min. If antibody staining was performed, fixation was followed by permeabilization in a 0.1% Triton X 100 solution. Samples were blocked in 5% fetal bovine serum in phosphate-buffered saline (blocking buffer) for 1 h. Samples were stained with primary antibody (1:100) in blocking buffer for 1 h followed by secondary antibody (1:100) and DAPI staining for 1 h. Images were taken on a Zeiss LSM710 confocal microscope using 63x or 100x objective lenses. For the mouse brain sections, additional images were captured on a BioTek Lionheart microscope at ×10 and image-stitched using the Biotek GEN5 v3.04 Imager software.
+ Open protocol
+ Expand
10

Immunofluorescence Staining of Activated Caspase-3

Check if the same lab product or an alternative is used in the 5 most similar protocols
Frozen sections (5 μm) were fixed with 4% paraformaldehyde-PBS 20 min at room temperature, permeabilized with 0.3% Triton X-100 in 1X PBS, blocked in 3% BSA (or 10% FBS) in 1X PBS, stained with primary antibodies for overnight at 4°C, followed by staining with 2° antibodies (1 hr) and DAPI stain. Samples were covered with coverglasses and imaged using Lionheart microscope (Biotek). Ac-caspase-3 monoclonal antibody (9661S, Cell Signaling, 1:200) was used to perform the immunofluorescence staining.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!