Permanox lab tek chamber slide

Manufactured by Thermo Fisher Scientific

Sourced in United States

Permanox Lab-Tek chamber slides are a type of cell culture vessel designed for microscopic analysis. These slides feature a removable chamber that allows for cell seeding, treatment, and observation under a microscope. The Permanox material provides a stable and reliable surface for cell attachment and growth.

Automatically generated - may contain errors

Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Fluoview fv10i confocal laser scanning microscope, by Olympus (1 mentions) Mitotracker deep red, by Thermo Fisher Scientific (1 mentions) S2 500, by Thermo Fisher Scientific (1 mentions) Tecnai g2 spirit transmission electron microscope, by Thermo Fisher Scientific (1 mentions) Tecnai g2 spirit, by Thermo Fisher Scientific (1 mentions) Em bed, by Electron Microscopy Sciences (1 mentions) Item imaging platform software, by Olympus (1 mentions) Libra 120, by Zeiss (1 mentions) Evos fl auto cell imaging system, by Thermo Fisher Scientific (1 mentions) Fluorescent mounting media, by Agilent Technologies (1 mentions) Cell f software, by Olympus (1 mentions) Vectashield plus dapi, by Vector Laboratories (1 mentions) Lsm 710 microscope, by Zeiss (1 mentions) Xarosa digital camera, by EMSIS (1 mentions)

11 protocols using permanox lab tek chamber slide

Microscopic Analysis of Macrophage Mitochondria

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

For electron microscopy, macrophages were seeded on 2-well Permanox Lab-Tek chamber slides (Nunc 177429). Macrophages were fixed as described[1 (link)]. Sample processing and imaging was performed at the Campus Microscopy and Imaging Facility at The Ohio State University as described[1 (link)]. Mitochondrial morphology was manually quantified using ImageJ software (National Institutes of Health, Bethesda, MD) as described[17 (link),18 (link)].
For fluorescence microscopy, macrophages were seeded on glass coverslips and stained with 250 nM MitoTracker Deep Red (Thermo Fisher Scientific M22426) for 15 minutes before infection. Macrophages were stained with 1 μg/mL Hoechst 33342 (Thermo Fisher Scientific 62249) for 15 min before fixation with 4% paraformaldehyde for 0.5 h. Images were captured using an Olympus Fluoview FV10i laser scanning confocal microscope with a 60x objective. MitoTracker was pseudocolored in red.

Hamilton K., Krause K., Badr A., Daily K., Estfanous S., Eltobgy M., Khweek A.A., Anne M.N., Carafice C., Baetzhold D., Tonniges J.R., Zhang X., Gavrilin M.A., Parinandi N.L, & Amer A.O. (2020). DEFECTIVE IMMUNOMETABOLISM PATHWAYS IN CYSTIC FIBROSIS MACROPHAGES. Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society, 20(4), 664-672.

+ Open protocol

+ Expand

Cerebellar Granule Neuron Culture and Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cerebellar granule neurons were plated at a density of 1 × 10⁵ cells/cm² on poly-L-lysine-coated, eight-well Permanox Lab-Tek chamber slides (Nunc) in Neurobasal-A medium supplemented with 2% (vol/vol) B27 (Gibco BRL), 0.5% (vol/vol) GlutaMAX, 100 U/ml penicillin and 100 μg/ml streptomycin. The CGN survival assay was performed as previously described
[14 (link)]. The neurite outgrowth assay was also performed as previously described
[15 (link)].

Klementiev B., Li S., Korshunova I., Dmytriyeva O., Pankratova S., Walmod P.S., Kjær L.K., Dahllöf M.S., Lundh M., Christensen D.P., Mandrup-Poulsen T., Bock E, & Berezin V. (2014). Anti-inflammatory properties of a novel peptide interleukin 1 receptor antagonist. Journal of Neuroinflammation, 11, 27.

+ Open protocol

+ Expand

Fixation and Imaging of Macrophages

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Macrophages were seeded in Permanox Lab-Tek chamber slides (177,429, Nunc) and then fixed with 2.5% glutaraldehyde (18,426, Ted Pella, Redding, CA, USA) in 0.1 M of phosphate buffer, pH 7.4 (S369 and S37, Fisher Scientific) containing 0.1 M of sucrose (S2-500, Fisher Scientific). Campus Microscopy & Imaging Facility at The Ohio State University performed sample processing as previously described (Krause et al., 2018a (link)). FEI Tecnai G2 Spirit transmission electron microscope plus AMT camera system was used for taking images.

Badr A., Eltobgy M., Krause K., Hamilton K., Estfanous S., Daily K.P., Abu Khweek A., Hegazi A., Anne M.N., Carafice C., Robledo-Avila F., Saqr Y., Zhang X., Bonfield T.L., Gavrilin M.A., Partida-Sanchez S., Seveau S., Cormet-Boyaka E, & Amer A.O. (2022). CFTR Modulators Restore Acidification of Autophago-Lysosomes and Bacterial Clearance in Cystic Fibrosis Macrophages. Frontiers in Cellular and Infection Microbiology, 12, 819554.

+ Open protocol

+ Expand

Macrophage Ultrastructure Visualization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Macrophages were cultured in 2-well Permanox Lab-Tek chamber slides (Nunc, 177,429) and fixed with 2.5% glutaraldehyde (Ted Pella, 18,426) in 0.1 M phosphate buffer, pH 7.4 (Fisher Scientific, S369 and S373) containing 0.1 M sucrose (Fisher Scientific, S2–500). Sample processing was performed by the Campus Microscopy & Imaging Facility at The Ohio State University as previously described [5]. Images were taken using a FEI Tecnai G2 Spirit transmission electron microscope plus AMT camera system.

Krause K., Caution K., Badr A., Hamilton K., Saleh A., Patel K., Seveau S., Hall-Stoodley L., Hegazi R., Zhang X., Gavrilin M.A, & Amer A.O. (2018). CASP4/caspase-11 promotes autophagosome formation in response to bacterial infection. Autophagy, 14(11), 1928-1942.

+ Open protocol

+ Expand

Ultrastructural Analysis of LNCAP Cells and Platelets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monolayer cell cultures of LNCAP cells and platelets were conducted in 8 wells Permanox Lab-Tek^® Chamber Slides (NUNC) and fixed at 4 °C in 1.5% glutaraldehyde, 1% formaldehyde, 0.05 M cocodilate buffer. Fixed cells were post-fixed in 1% osmium tetroxide for 1 hour at 4 °C, washed in distilled water, and treated with 0.15% tannic acid and 2% uranyl acetate. Then, dehydration through graded alcohols and propylene oxide, and then EMbedding in EMbed (Electron Microscopy Sciences) was done. Ultrathin sections (50-70 nm) were stained with 1% uranyl acetate and lead citrate 22 (link). Samples were prepared and examined in a Transmission Electron Microscope using the Libra 120 (Zeiss) ITEM Imaging Platform Software (Olympus) at the Centre for Scientific Instrumentation (CIC) of the University of Granada.

Rodriguez-Martinez A., Simon-Saez I., Perales S., Garrido-Navas C., Russo A., de Miguel-Perez D., Puche-Sanz I., Alaminos C., Ceron J., Lorente J.A., Molina M.P., Gonzalez C., Cristofanilli M., Ortigosa-Palomo A., Real P.J., Rolfo C, & Serrano M.J. (2022). Exchange of cellular components between platelets and tumor cells: impact on tumor cells behavior. Theranostics, 12(5), 2150-2161.

+ Open protocol

+ Expand

Measuring Cellular Oxidative Stress

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cellular ROS production was measured by fluorescence probe dihydroethidium (DHE) which allows intracellular detection of superoxide radicals (O 2 .-). Cells were seeded 24 h before radiation exposure in 96-well tissue culture plates at concentration of 1 × 10 5 cells/ml. After irradiation cells were washed with phosphate buffer saline (PBS, Sigma) and incubated with 20 μM DHE for 30 min at 37°C. Fluorescence intensity was quantified by means of plate reader device at an excitation wavelength of 485 nm and emission wavelength of 535 nm. The image analysis of intracellular ROS was carried out by seeding the cells on Permanox Lab-Tek Chamber Slides (Nunc Naperville, USA) at concentration of 2.5 × 10 4 cells/ml. The next day, after radiation exposure, cell culture medium was removed and samples were washed with PBS. Non-irradiated and positive control cell samples exposed to H 2 O 2 for 15 min were also included. After the treatment cells were incubated with 20 μM DHE (Sigma) for 30 min in the dark. At the end of incubation period fluorescent dye was removed and cells were washed four times with PBS. Fluorescence of the cells was examined by EVOS FL Auto Cell Imaging System (ThermoFisher Scientific, Waltham, USA) using 585 nm excitation and 624 nm emission filter.

Marjanovic Cermak A.M., Pavicic I., Tariba Lovakovic B., Pizent A, & Trosic I. (2017). In vitro non-thermal oxidative stress response after 1800 MHz radiofrequency radiation. General physiology and biophysics, 36(4).

+ Open protocol

+ Expand

Evaluating App Invasion of Endothelial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

To test App invasion, endothelial cells were seeded (10⁴) (37 (link)) into permanox Lab-Tek chamber slides (Thermo Scientific) and incubated overnight. Cells were infected with the different App strains with a MOI of 3:1. After a 3-h incubation, non-adherent bacteria were removed by a DPBS wash and subsequently fixed 30 min using formaldehyde 3.7% at room temperature. Then cells were gently wasehed three times with DPBS.
After fixation, endothelial cells' membrane was permeabilized with 0.1% Triton X-100 (US Biological, Salem, Ma, USA) for 5 min at room temperature (43 (link), 44 (link)), followed by 2 washes with DPBS. Once the membrane was permeabilized to observe the implication of different cellular organelles in App invasion by immunofluorescent labeling different labels were used according to the organelle to be observed, which are described below. Several repetitions were performed for each organelle.

Plasencia-Muñoz B., Avelar-González F.J., De la Garza M., Jacques M., Moreno-Flores A, & Guerrero-Barrera A.L. (2020). Actinobacillus pleuropneumoniae Interaction With Swine Endothelial Cells. Frontiers in Veterinary Science, 7, 569370.

+ Open protocol

+ Expand

Live/Dead Cell Viability Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

The LIVE/DEAD viability kit for mammalian cells (Invitrogen, ThermoFisher) was used to simultaneously stain live (green) and dead (red) cells after injury was performed. hCEC were seeded in 8-well Permanox Lab-Tek chamber slides (ThermoFisher Scientific). Cell monolayers were incubated with a solution containing 2 µmol/L calcein AM and 4 µmol/L ethidium homodimer-1 in PBS for 30 min. After incubation, the staining solution was discarded, chambers removed, and slides mounted under coverslips in fluorescent mounting media (Dako, United Kingdom). Imaging was performed using an upright fluorescence microscope (BX51, Olympus, Southend-on-Sea, United Kingdom) with images captured with a black and white camera (XM-10, Olympus) and Cell^F software (Olympus).

Orozco Morales M.L., Marsit N.M., McIntosh O.D., Hopkinson A, & Sidney L.E. (2019). Anti-inflammatory potential of human corneal stroma-derived stem cells determined by a novel in vitro corneal epithelial injury model. World Journal of Stem Cells, 11(2), 84-99.

+ Open protocol

+ Expand

Evaluating Sporozoite Invasion and Liver Cell Development

Check if the same lab product or an alternative is used in the 5 most similar protocols

EXAMPLE 2

Functional Assay 1: Liver Cell Invasion Assay

Invasion assay was performed as described by Sattabongkot, J. et al. (2006, Am. J. Trop. Med. Hyg. 74, 708-715. HC-04 monolayers were prepared by seeding ˜60,000 cells into individual wells of Permanox Labtek chamber slides (Thermo Fisher Scientific, Waltham, Mass.). Sporozoites were concentrated by centrifugation and 30,000-80,000 sporozoites (depending on replica) were applied to monolayer; PBS was applied to at least one well per replica as a negative control. After a 3-hr incubation, free sporozoites were removed by washing 3 times with complete media. Slides were incubated at 37° C. with 5% CO₂and media was changed daily. On day 5-6, slides were fixed with cold methanol, blocked with BSA and stained with anti-HSP-70 as the primary antibody and AlexaFluor488 anti-IgG as the secondary.

Slides were mounted using Vectashield plus DAPI (Vector Laboratories, Burlingame, Calif.). Growth of sporozoites into exo-erythrocytic forms was observed using epifluorescent microscopy in 5 biological replicates.

Our results showed that two different cultured sporozoites uniformly express circumsporozoite protein (CSP) on their surface and that parasites are capable of invading and developing inside hepatic cells (HC-04 cells, two different replicate cultures on day 6) (data not shown).

US10577578B2. Method for developing malaria sporozoites in vitro (2020-03-03). WALTER REED ARMY INSTITUTE OF RESEARCH DEPARTMENT OF THE ARMY [US], None [US], None [US]. Inventors: Tatyana Savransky [US], Jackie Williams [US].

+ Open protocol

+ Expand

Visualizing GSDMB Protein Localization

Check if the same lab product or an alternative is used in the 5 most similar protocols

GSDMB constructs were visualized by immunofluorescence in transient transfected (72 h) and 4% paraformaldehyde-fixed cells with the indicated primary antibodies (Supplementary Table 2) as described before [42 (link)]. Confocal microscopy images were captured by LSM710 microscope (Zeiss) and processed by Fiji software (Image J 1.52). Alternatively, live cell tracking imaging was performed with doxycycline inducible vectors expressing GFP-tagged GSDMB constructs (Supplementary information). To evaluate mitochondrial morphology correlative light and electron microscopy (CLEM) procedures were performed in 23132/87 cells. Cells were seeded in a permanox Lab-Tek chamber slide (Nalge Nunc International) and transfected with doxycycline inducible vectors. After 6 h of transfection, cells were induced with Doxycycline at 200 ng/ml, incubated with red MitoTracker^TM Deep Red FM (ThermoFisher) for 30 min at 37 °C and fixed in 3% glutaraldehyde. The confocal images were acquired with a Leica TCS SP8 HyVolution II (Leica Microsystems). After fluorescence capture, slides were processed for transmission electron microscopy analysis with FEI Tecnai Spirit BioTwin (ThermoFisher). Pictures were taken using Radius software (Version 2.1) with a Xarosa digital camera (EMSIS GmbH).

Oltra S.S., Colomo S., Sin L., Pérez-López M., Lázaro S., Molina-Crespo A., Choi K.H., Ros-Pardo D., Martínez L., Morales S., González-Paramos C., Orantes A., Soriano M., Hernández A., Lluch A., Rojo F., Albanell J., Gómez-Puertas P., Ko J.K., Sarrió D, & Moreno-Bueno G. (2023). Distinct GSDMB protein isoforms and protease cleavage processes differentially control pyroptotic cell death and mitochondrial damage in cancer cells. Cell Death and Differentiation, 30(5), 1366-1381.

+ 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?

Revolutionizing how scientists
search and build protocols!