Log In Sign up
The largest database of trusted experimental protocols

Sp8 confocal system

Manufactured by Leica camera
Visit Supplier
Sourced in Germany

The Leica SP8 confocal system is a high-performance imaging platform designed for advanced microscopy applications. It utilizes the principles of confocal laser scanning microscopy to capture detailed, high-resolution images of biological samples. The SP8 system is equipped with multiple laser lines, sensitive detectors, and a versatile optical configuration, enabling researchers to obtain comprehensive data from their specimens.

Automatically generated - may contain errors

Lab products found in correlation

Rhodamine phalloidin, by Merck Group (3 mentions) Sp8 laser scanning confocal system, by Leica camera (2 mentions) Goat anti mouse alexa 488, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Fluo 4 am, by Thermo Fisher Scientific (2 mentions) Lsm 780, by Zeiss (2 mentions) T8787, by Merck Group (1 mentions) Anti p65 antibody c 20, by Santa Cruz Biotechnology (1 mentions) Goat serum, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Santa Cruz Biotechnology (1 mentions) Image it fixation permeabilization kit, by Thermo Fisher Scientific (1 mentions) Alexa fluor secondary antibody, by Jackson ImmunoResearch (1 mentions) Stellaris 8 confocal microscope, by Leica camera (1 mentions) Mouse anti yap, by Santa Cruz Biotechnology (1 mentions) Mouse anti β1 integrin, by Abcam (1 mentions) Rabbit anti paxillin, by Abcam (1 mentions) Rabbit anti p fak, by Abcam (1 mentions) Mouse anti runx2, by Abcam (1 mentions) Sp8 confocal microscope, by Leica camera (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

47 protocols using sp8 confocal system

1

Immunofluorescence Analysis of HT-29 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunofluorescence analysis, HT-29 cells were grown on coverslips. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% TritonX-100 (Sigma-Aldrich, T8787) and blocked with 5% bovine serum albumin (BSA). Blocked cells were incubated with indicated primary antibody overnight at 4 °C. Cells were then incubated with secondary antibody for 1 h at room temperature. Fluorescence imaging was done on Leica SP8 Confocal System.
Zu R., Yu Z., Zhao J., Lu X., Liang W., Sun L., Si C., Zhu K., Zhang T., Li G., Zhang M., Zhang Y., Liu N., Yuan J, & Shan B. (2021). Quantitative analysis of phosphoproteome in necroptosis reveals a role of TRIM28 phosphorylation in promoting necroptosis-induced cytokine production. Cell Death & Disease, 12(11), 994.
+ Open protocol
+ Expand
2

Immunofluorescence Analysis of p65 in HT-29 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
HT-29 cells were plated on coverslips and treated as indicated. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100 in PBS. Cells were then incubated with 5% bovine serum albumin in PBS for 30 min and the anti-p65 antibody (C-20, Cat# sc-372, Santa Cruz) overnight at 4 °C, followed by three PBS washes and subsequent incubation with the fluorescent secondary antibody for 1 h at room temperature. DAPI (300 nM) (Thermo Fisher Scientific) was included in the final wash for nuclei counterstain. Cells were mounted with 10% glycerol and images were acquired with a laser scanning microscope (Leica SP8 Confocal System).
Zhu K., Liang W., Ma Z., Xu D., Cao S., Lu X., Liu N., Shan B., Qian L, & Yuan J. (2018). Necroptosis promotes cell-autonomous activation of proinflammatory cytokine gene expression. Cell Death & Disease, 9(5), 500.
+ Open protocol
+ Expand
3

Imaging Activated NK Cell Adhesion

Check if the same lab product or an alternative is used in the 5 most similar protocols
IL2-activated human NK cells were attached in the microfluidic channel as described above and exposed to C3a-containing medium (2 μg/mL mixed with migration medium, 0.4% BSA in RPMI) or IgG control for 20 minutes. The cells were fixed [PFA 4% in PBS (Santa Cruz Biotechnology, Cat# 30525–89-4)], washed, and blocked with 10% goat serum (Thermofisher Scientific, Cat# 31872) for 45 minutes at room temperature. Primary rabbit anti-human CD11a (Cat#ab52895, Abcam, USA), primary rabbit anti-human CD18 (phospo-T758; Cat#ab63388, Abcam, USA), primary mouse anti-human CD11a+CD18 antibody [24] (Cat# ab13219, Abcam, USA), or respective control antibody (Rabbit IgG and Mouse IgG1) was diluted as suggested by the provider, infused into the microfluidic devices, and incubated for overnight at 4oC. Samples were then washed three times and incubated with diluted secondary antibody (1:200 dilution; anti-mouse goat IgG, Cat# A32723 or anti-rabbit goat IgG, Cat# A32721, Thermofisher Scientific, USA) in the dark at room temperature for one hour. The samples were washed three times and exposed to DAPI in the dark at room temperature for 10 minutes. After washing, samples were preserved at 4˚C for confocal microscopy analysis. Images were taken using Leica SP8 confocal system at 10X object (zoom 5).
Nandagopal S., Li C.G., Xu Y., Sodji Q.H., Graves E.E, & Giaccia A.J. (2021). C3aR Signaling Inhibits NK-cell Infiltration into the Tumor Microenvironment in Mouse Models. Cancer immunology research, 10(2), 245-258.
+ Open protocol
+ Expand
4

Immunofluorescence Imaging of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were fixed and permeabilized with Image-iT® Fixation/Permeabilization Kit (Thermo Fisher Scientific) and were blocked in blocking buffer (0.1 M glycine, 2% BSA, 0.1% Triton-X100 in PBS). Primary antibodies were diluted in blocking buffer and incubated at 4 °C overnight. After washing, samples were incubated with alexa488-, alexa568- and alexa647-conjugated secondary antibodies for 2 h at room temperature and thereafter mounted using Dako mounting media. Samples were captured using an SP8 confocal system (Leica, Wetzlar, Germany) and image analysis was performed using Image J (NIH, USA)
Gao H., Zheng W., Li C, & Xu H. (2021). Isoform-Specific Effects of Apolipoprotein E on Hydrogen Peroxide-Induced Apoptosis in Human Induced Pluripotent Stem Cell (iPSC)-Derived Cortical Neurons. International Journal of Molecular Sciences, 22(21), 11582.
+ Open protocol
+ Expand
5

Immunofluorescence Staining and Confocal Imaging

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were fixed for 30 min with 4% paraformaldehyde (PFA), washed with 1× phosphate-buffered saline (PBS), and permeabilized with 0.5% Triton X-100/1× PBS for 30 min, blocked with 10% donkey serum for 1 h, and incubated with mouse anti-FLAG monoclonal or anti-2A primary antibodies in 0.1% Triton X-100/1× PBS overnight at 4°C. The next day, cells were washed with 1× PBS and incubated with the corresponding Alexa Fluor secondary antibodies (Jackson ImmunoResearch Laboratories; donkey anti-rabbit, anti-mouse, IgG (H+L) 488, 594, or 680) at a 1:400 dilution for 1 h at room temperature, using Hoechst 33258 (1:5,000) as a nuclear counterstain. Fluorescent confocal images were acquired and analyzed using the Leica SP8 confocal system.
Zhu Y., Saribas A.S., Liu J., Lin Y., Bodnar B., Zhao R., Guo Q., Ting J., Wei Z., Ellis A., Li F., Wang X., Yang X., Wang H., Ho W.Z., Yang L, & Hu W. (2023). Protein expression/secretion boost by a novel unique 21-mer cis-regulatory motif (Exin21) via mRNA stabilization. Molecular Therapy, 31(4), 1136-1158.
+ Open protocol
+ Expand
6

Confocal Microscopy and FRET Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Confocal images were captured by a Leica SP8 laser scanning confocal system with a 63 × water lens or Leica STELLARIS 8 confocal microscope with a 93 × glycerol lens following manufacturer’s instructions. FRET analysis was conducted on the same Leica SP8 confocal system according to the manufacturer’s instruction using 405 nm and 514 nm lasers. For surface rendering of ER sheets, Imaris 9.8.0 (Bitplane) was used as previously described (63 (link)).
Li B., Niu F., Zeng Y., Tse M.K., Deng C., Hong L., Gao S., Lo S.W., Cao W., Huang S., Dagdas Y, & Jiang L. (2023). Ufmylation reconciles salt stress-induced unfolded protein responses via ER-phagy in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 120(5), e2208351120.
+ Open protocol
+ Expand
7

Quantifying Retinal Immunohistochemistry Signals

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunohistochemistry signals in wholemount retinas were analyzed using SP8 Confocal system (LEICA, Wetzlar, Germany). Signals were assessed from 1024 × 1024-pixel z-stack images at 0.5 μm z steps taken with 63× glycerol or 40× oil immersion lens. Images were processed using Fiji software, for quantification of fluorescence intensity profile images were converted to 8-bit type then intensity threshold was measured in black and white.
Salman A., Hutton S.B., Newall T., Scott J.A., Griffiths H.L., Lee H., Gomez-Nicola D., Lotery A.J, & Self J.E. (2020). Characterization of the Frmd7 Knock-Out Mice Generated by the EUCOMM/COMP Repository as a Model for Idiopathic Infantile Nystagmus (IIN). Genes, 11(10), 1157.
+ Open protocol
+ Expand
8

Immunofluorescence Staining of Integrin Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were fixed using 4% paraformaldehyde/PBS for 15 min at room temperature, washed three times with washing buffer (0.1% Tween-20/PBS, 5 min), and permeabilized with 1% Triton X-100/PBS for 30 min. Samples were incubated in blocking buffer (3% BSA, 2% goat serum in PBS) for 30 min, then incubated with various primary antibodies including mouse anti-YAP (Santa Cruz Biotechnology, sc-101199), mouse anti-αVβ3-integrin (Abcam ab78614), mouse anti-β1-integrin (Abcam ab24693), rabbit anti-paxillin (Abcam ab32084), rabbit anti-pFAK (Abcam ab81298), mouse anti-RUNX2 (Abcam ab76956) overnight at 4 °C on a shaker. All antibodies were diluted in buffer at 1:100 except YAP antibody was diluted at 1:300. After washing, samples were incubated with corresponding secondary antibodies including Alexa 488 Goat-anti-mouse (Invitrogen A11001), rhodamine goat-anti-rabbit (Millipore AP132), rhodamine-phalloidin (Sigma P1951) for 1 h at room temperature on a shaker. All secondary antibodies were diluted at 1:300. Cell nucleus counter stain was performed using Hoeschst nuclear stain (Cell Signaling Technology 4082S, 2 ug/mL). Samples were washed with washing buffer (three times, 5 min per wash) before being imaged using a confocal microscope (40x oil immersion, Leica SP8 confocal system). All images were processed using open-source Fiji software [35 (link), 36 (link)].
Stanton A.E., Tong X., Lee S, & Yang F. (2019). Biochemical Ligand Density Regulates YAP Translocation in Stem Cells through Cytoskeletal Tension and Integrins. ACS applied materials & interfaces, 11(9), 8849-8857.
+ Open protocol
+ Expand
9

Carotid Atherosclerosis Model in ApoE-deficient Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
ApoE genetically defective C57BL/6 mice were prepared to build carotid atherosclerosis model. After carotid artery ligation and high‐fat diet for one month, a mouse model of carotid atherosclerosis was build. The carotid artery was removed and frozen sections were prepared, oil red O staining was used to analyze the formation of atherosclerotic plaque. After the carotid atherosclerosis model is successfully constructed, the carotid artery was exposed, probe‐loaded hydrogel was applied on the surface of blood vessel, incubated for 10 min, then the probe‐loaded hydrogel was removed by filter paper adsorption, then the fluorescence of the blood vessel position was continuously observed and photographed under the living imaging system for 1 h. Then carotid arteries were removed for imaging. After that, the blood vessels were fixed with 4% paraformaldehyde for 24 h, dehydrated with 40% sucrose for 3 d, and then frozen sectioned. Fluorescence was observed in Leica SP8 confocal system.
Sang M., Huang Y., Wang L., Chen L., N.a.w.s.h.e.r.w.a.n., Li G., Wang Y., Yu X., Dai C, & Zheng J. (2023). An “AND” Molecular Logic Gate as a Super‐Enhancers for De Novo Designing Activatable Probe and Its Application in Atherosclerosis Imaging. Advanced Science, 10(12), 2207066.
+ Open protocol
+ Expand
10

Cytosolic Calcium Imaging of Alpha-Synuclein Fibrils

Check if the same lab product or an alternative is used in the 5 most similar protocols
For cytosolic calcium measurements, cells were loaded with the Ca2+ indicator Fluo-4/AM (Thermo Fisher Scientific) for 15 min at 37ºC, placed onto the stage of a Leica SP8 Confocal System and continuously perfused with HEPES buffer solution or HEPES Ca2+-free buffer (HEPES buffer solution: 142.2 mM NaCl, 5.4 mM KCl, 1.0 mM NaH2PO4, 10 mM HEPES, 5.6 mM dextrose, 0.8 mM MgSO4 and 1 mM CaCl2/ HEPES Ca2+-free buffer: 142.2 mM NaCl, 5.4 mM KCl, 1.0 mM NaH2PO4, 10 mM HEPES, 5.6 mM dextrose, 0.8 mM MgSO4 and 1 mM EGTA). 1 µM αSyn PFFs was used to trigger Ca2+ release. Data are expressed as fluorescence/baseline fluorescence × 100% of the average values of samples from 3 biological replicates (at least 25 cells were individually analyzed). The images were obtained using a Leica SP8 confocal microscope, using a × 63 objective lens, 1.4 NA, excitation at 488 nm and emission at 505–525 nm for both dyes.
Rodrigues P.V., de Godoy J.V., Bosque B.P., Amorim Neto D.P., Tostes K., Palameta S., Garcia-Rosa S., Tonoli C.C., de Carvalho H.F, & de Castro Fonseca M. (2022). Transcellular propagation of fibrillar α-synuclein from enteroendocrine to neuronal cells requires cell-to-cell contact and is Rab35-dependent. Scientific Reports, 12, 4168.
+ 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!