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Lipofuscin

Lipofuscin is a pigment that accumulates in various cell types, particularly in postmitotic cells such as neurons and cardiomyocytes, as a byproduct of normal cellular metabolism.
It is considered a hallmark of aging and is associated with a number of age-related diseases.
Lipofuscin is composed of cross-linked proteins, lipids, and metals, and its accumulation can impair cellular function and contribute to the development of pathological conditions.
Researchers studying lipofuscin can optimize their efforts with PubCompare.ai, an AI-powered tool that enhances reproducibility and accuracy by easily locating the best protocols from literature, pre-prints, and patents using AI-driven comparisons.
This streamlines the research process and helps researchers get the most accurate results.

Most cited protocols related to «Lipofuscin»

1
Optimized Lipofuscin Visualization by SBB Staining
Cited 20 times
Combining the protocols of Gatenby et al. [17 (link)], and Rasmussen, [18 (link)] for SBB staining, we achieved optimal lipofuscin visualization in cell cultures and tissues with the following methodology:

Preparation of SBB solution: 0.7gr of SBB (BDH, Vizas, Athens, Greece) was dissolved in 70% ethanol, covered with parafilm and thoroughly stirred overnight at room temperature. Filtered through filter paper and then filtered again through frittered glass filter of medium porosity with suction. Throughout the process, it was important to avoid ethanol evaporation,which results in precipitation of the stain, so the solution was storedin an airtightcontainer.

Staining Procedure: OCT-Frozen-sections mounted onto superfrost slides were fixed in 1% (wt/vol) formaldehyde/PBS for 1 min at room temperature and then washed three times (approx.1 min) at room temperature, with PBS. Sections were then incubated for 5 min in 50% ethanol and then for another 5 min into 70% ethanol. Coverslips with fixed cells were incubated for 2 min in 70% ethanol. Tissue samples were dewaxed with xylene and dehydrated until 70% ethanol. In order to avoid precipitation of SBB on cells or tissues the following two steps are crucial: 1) a drop from freshly prepared SBB was dropped on a clean slide. The coverslip with the cells or the dehydrated tissue on a slide was placed facing down on the drop of SBB on the slide. The staining was observed under the microscope. The desirable outcome with no precipitation was accomplished by 2-8 minutes. 2) The coverslip or the slide, were carefully lifted and the SBB on the edges of the coverslip or the tissue-slide was wiped out manually from the back and along the edges of the coverslip or the slide with the help of a soft paper. The cells or the tissues were then embedded into 50% ethanol, transferred and washed in distilled water, counterstained with 0.1% Nuclear Fast Red (NFR) (Sigma, BioLine, Athens Greece) for 10 min., and mounted into 40% Glycerol/TBS mounting medium. Lipofuscin staining was considered positive when perinuclear and cytoplasmic aggregates of blue-black granules were evident inside the cells.

Georgakopoulou E., Tsimaratou K., Evangelou K., Fernandez M.P., Zoumpourlis V., Trougakos I., Kletsas D., Bartek J., Serrano M, & Gorgoulis V. (2012). Specific lipofuscin staining as a novel biomarker to detect replicative and stress-induced senescence. A method applicable in cryo-preserved and archival tissues. Aging (Albany NY), 5(1), 37-50.
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Publication 2012
Cell Culture Techniques Cells Cytoplasmic Granules Ethanol Formaldehyde Frozen Sections Glycerin Lipofuscin Microscopy Stains Suction Drainage Tissues Xylene
2
Ex Vivo Imaging of Neovascular AMD Tissue
Cited 13 times

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Publication 2015
4-phenylenediamine Allium cepa ARID1A protein, human Autopsy Blood Vessel Bruch Membrane Buffers Choroid Donors Edema Epoxy Resins Eye Glutaral Immersion Lens, Crystalline Light Lipids Lipofuscin Macula Lutea Melanosomes Microscopy Multimodal Imaging Optic Disk Osmium paraform Pathologic Neovascularization Phosphates Photoreceptor Cells Pigmentation Polybed 812 Radionuclide Imaging Retina Tannins Tissue, Membrane Tissues Tolonium Chloride Woman
3
Longitudinal Imaging of Stem Cell Co-Culture
Cited 13 times

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Publication 2012
Afterimage Coculture Techniques Fluorescence Lipofuscin Microscopy Microscopy, Confocal Microscopy, Fluorescence NADH PYCARD protein, human Sapphire Stainless Steel Submersion Tissues Titanium
4
Multiplexed in situ RNA detection
Cited 11 times

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Publication 2017
2',5'-oligoadenylate Acid Hybridizations, Nucleic Biopsy Ethanol Fluorescence Frozen Sections Genes Lipofuscin Neoplasms Oligonucleotide Primers Oligonucleotides Reverse Transcription Tissues
5
Lifespan and Stress Assays in C. elegans
Cited 11 times
For aging assays, synchronous populations were obtained by allowing 5–10 hermaphrodites to lay eggs for 4 h to overnight. For ease of analysis, we used the adult sterile strain, fem-1(hc17), as the wild-type strain to avoid progeny overgrowth in lifespan assays. Therefore, eggs were shifted to 25 °C, the nonpermissive temperature for fertility of fem-1(hc17). Lifespan scoring was initiated after hermaphrodites completed the final larval molt, on the first day of adulthood. For aging assays with BB extracts, treatments were added to NGM agar plates on the first day of the lifespan assay. For lifespan assays with fertile strains, hermaphrodites were transferred daily for the first 4 days of adulthood to avoid progeny overgrowth. In these cases, all treatment plates were prepared on day 0 of adulthood. Statistical analyses and survival plots of lifespan data were performed with JMP analysis software (SAS Institute Inc, Cary, NC, USA). Pharynx pumping rates were scored on adults at room temperature (24 °C) under a Nikon SMZ1500 stereomicroscope (Nikon, Melville, NY, USA).
Thermotolerance assays were performed with hermaphrodites on adult day 5, after the majority of egg-laying had ceased. Animals were transferred onto 3-cm NGM agar plates supplemented as indicated and then incubated at 35 °C for 16 h. Survival was scored as the number of animals responsive to gentle touch as a fraction of the original number of animals on the plate. Animals that had died from dessication on the sides of the plate were censored. Paraquat-induced oxidative stress assays were performed with fem-1(hc17) hermaphrodites at 25 °C as for aging assays, except paraquat was added to NGM medium to 10 mm final concentration (ChemService, West Chester, PA, USA). For hydrogen peroxide, we scored 5-h survival of adult day 5 fem-1(hc17) animals in S-basal medium with indicated concentrations of hydrogen peroxide.
To determine lipofuscin levels, adult hermaphrodites were anesthetized in 0.2% sodium azide and mounted on 2% agarose pads for visualization of intestinal fluorescence on a Nikon E800 microscope using an Endow GFP filter with a mercury UV source (Nikon). Images were captured using a constant exposure time with a Hamamatsu ORCA digital CCD camera (Hamamatsu, Bridgewater, NJ, USA) using OpenLab software (Improvision, Lexingon, MA, USA). Lipofuscin levels were measured using ImageJ software (NIH Image) by determining average pixel intensity in each animal's intestine.
To measure levels of 4-HNE, animals were collected, fixed with 4% formaldehyde and permeabilized by digestion with type IV collagenase (Sigma Chemical Company), as described (Loer & Kenyon, 1993 (link)). Fixed and permeabilized specimens were incubated with anti4-HNE antisera (1 : 100 dilution, Genox Corp, Baltimore, MD, USA) for 2 h at 24 °C, washed and incubated overnight at 4 °C with Alexafluor-546-conjugated goat anti-mouse secondary antibody (1 : 100) (#A-11003, Invitrogen, Carlsbad, CA, USA). Stained animals were mounted on 2% agarose pads and fluorescence visualized as for lipofuscin with appropriate filter sets. 4-HNE immunofluorescence was measured in the pharynx terminal bulb and somatic gonad using ImageJ software.
Wilson M.A., Shukitt-Hale B., Kalt W., Ingram D.K., Joseph J.A, & Wolkow C.A. (2006). Blueberry polyphenols increase lifespan and thermotolerance in Caenorhabditis elegans. Aging Cell, 5(1), 59-68.
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Publication 2006
Adult Agar Animals Antibodies, Anti-Idiotypic Biological Assay Desiccation Digestion Diploid Cell Eggs Fertility Fingers Fluorescence Fluorescent Antibody Technique Formaldehyde Goat Gonads Hermaphroditism Immune Sera Intestines Larva Lipofuscin Matrix Metalloproteinase 2 Mercury Mice, House Microscopy Molting Orcinus orca Oxidative Stress Paraquat Peroxide, Hydrogen Pharynx Plant Bulb Population Group Sepharose Sodium Azide Sterility, Reproductive Strains Technique, Dilution Thermotolerance Touch

Most recents protocols related to «Lipofuscin»

1
Kir4.1 Immunohistochemistry in EAE and MS
Cryostat sections prepared from 4% paraformaldehyde-perfused control or MOG35-55-induced EAE female mice were subjected to heat-induced epitope retrieval (HIER) and incubated overnight at 4°C with anti-e1 serum and rabbit anti-Cter Kir4.1356-375 antibody, revealed by AF594-coupled and AF488-coupled secondary antibodies, respectively. Sections were stained with DAPI and coverslipped with anti-fading mounting medium, and pictures were taken at fixed fluorescence exposure. Peptide-N-glycosidase F (PNGase F, New England BioLabs) was used to evaluate the effect of N-linked glycosylation on the anti-e1 reactivities. For this, HIER-treated sections were incubated with 5 U/µl of PNGase F in 10 mM PBS, 10 mM EDTA at pH 7.6 at 37°C overnight, before being processed for immunohistofluorescence.
For fresh-frozen human tissues, 12µm-thick cryostat sections enriched in subcortical WM (Supplementary Table 3) were prepared from selected blocks containing inflamed subcortical WM or NAWM,35 (link) defined from CD68 and Luxol Fast Blue stainings (Supplementary Table 4). Acetone-fixed sections were processed for Kir4.1 immunostaining as described above, and incubated with 0.1% Black Soudan to stain the white matter and hide lipofuscin-driven autofluorescence before covering with anti-fade mounting medium. For quantification, three fields at ×40 objective of subcortical WM per sample were acquired at fixed fluorescence exposure time, and the average level of Kir4.1 immunofluorescence was measured with ImageJ software. Sections from four human renal fresh-frozen biopsies were also used for Kir4.1 immunofluorescence: control cortical pre-implant biopsies from two donors and cortical kidney biopsies with chronic inflammation from two patients with interstitial fibrosis/tubular atrophy.
Nicot A.B., Harb J., Garcia A., Guillot F., Mai H.L., Mathé C.V., Morille J., Vallino A., Dugast E., Shah S.P., Lefrère F., Moyon M., Wiertlewski S., Le Berre L., Renaudin K., Soulillou J.P., van Pesch V., Brouard S., Berthelot L, & Laplaud D.A. (2023). Aglycosylated extracellular loop of inwardly rectifying potassium channel 4.1 (KCNJ10) provides a target for autoimmune neuroinflammation. Brain Communications, 5(2), fcad044.
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Publication 2023
Acetone Antibodies Antibodies, Anti-Idiotypic Atrophy Biopsy DAPI Donors Edetic Acid Endo-beta-N-Acetylglucosaminidase F Epitopes Females Fibrosis Fluorescence Fluorescent Antibody Technique Freezing Frozen Sections Glycopeptidase F Homo sapiens Kidney Kidney Cortex Lipofuscin Luxol Fast Blue MBS Mus Nephritis paraform Patients Protein Glycosylation Rabbits Serum Staining Stains Tissues White Matter
2
Moesin Immunofluorescence in Brain Tissue
For Moesin immunofluorescence, frozen pieces of brain tissue from temporal cortex were sectioned at -20 ˚C and transferred to microscope slides. Samples were then warmed to room temperature and immediately incubated in 4% PFA at room temperature for 10 minutes. Slides were then rinsed in diH2O and immersed in sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0) and incubated above a 240 W LED light source (HTG Supply, Cat. No. LED-6B240) at 4°C for four hours to reduce lipofuscin autofluorescence. Next, slides were incubated in blocking solution (2% non-fat milk in PBS plus 0.3% TritonX (PBSTr)) at 4°C for 30 minutes. Following non-specific blocking, slides were incubated overnight in blocking solution containing primary antibodies. The following day, slides were rinsed three times in PBSTr and incubated in blocking solution containing secondary antibodies at room temperature for one hour. Next slides were rinse three times using PBSTr, mounted with DAPI containing media, and coverslipped. Brains were visualized by confocal microscopy (Zeiss LSM 780 NLO with Examiner, Zeiss LSM 810 with Airyscan), and ImageJ.111 (link) Immunofluorescence was quantified by measuring average Moesin signal intensity within the nucleus of 50 neurons per biological replicate. For each sample, images were converted to 8-bit binary Z-projections using the Max Intensity projection setting and thresholded with the default parameters in ImageJ. Total fluorescence for each biological replicate was calculated by taking the product of the mean gray value and percent area for each of the 50 regions of interest selected and averaged. Antibodies, reagents, concentrations, and sources are listed in Table S11.
Beckmann A., Ramirez P., Gamez M., Gonzalez E., De Mange J., Bieniek K.F., Ray W.J, & Frost B. (2023). Moesin is an effector of tau-induced actin overstabilization, cell cycle activation, and neurotoxicity in Alzheimer’s disease. iScience, 26(3), 106152.
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Publication 2023
Antibodies Biopharmaceuticals Brain Buffers Cell Nucleus DAPI DNA Replication Fluorescence Fluorescent Antibody Technique Freezing Light Lipofuscin Microscopy Microscopy, Confocal Milk, Cow's MSN protein, human Neurons Sodium Citrate Temporal Lobe Tissues Tween 20
3
Quantifying Hippocampal Tau Pathology in Brain Tissue
Formalin-fixed, paraffin-embedded tissue sections were cut at 5 μm from the mid-hippocampus at the level of the lateral geniculate nucleus. Tissue sections were deparaffinized and citrate antigen retrieval (ImmunoRetriever with citrate, Bio SB Inc, Goleta, CA) was performed at high heat and high pressure for 15 minutes (TintoRetriever Pressure Cooker, Bio SB Inc, Goleta, CA). Sections were labeled first with antibodies against synaptophysin and NeuN, then with secondary antibodies Alexa 488 goat anti-mouse IgG1 (1:400, A21121, Invitrogen Thermo Fisher Scientific, Waltham, MA) and Alexa 647 goat anti-rabbit (1:400, A21245, Invitrogen Thermo Fisher Scientific, Waltham, MA), followed by a 1-hour incubation with the labeled AT8 antibody. Autofluorescence was quenched using TrueBlack Plus Lipofuscin Autofluorescence Quencher (Biotium, Fremont, CA) diluted 1:40 in 70% ethanol and applied for 50 seconds. Hippocampal regions were marked based on anatomic landmarks by a Board-certified neuropathologist. Images were taken at 400X on a Zeiss LSM 880 confocal microscope (Zeiss, Oberkochen, Germany), with 1–3 images taken per region per case in the molecular layer of the dentate gyrus, stratum radiatum of CA1 and CA2, and the stratum radiatum/lucidum of CA3. Image acquisition parameters were uniform across the entire cohort. Experimenters were blinded during imaging and processing.
Morris M., Coste G.I., Redding-Ochoa J., Guo H., Graves A.R., Troncoso J.C, & Huganir R.L. (2023). Hippocampal Synaptic Alterations Associated with Tau Pathology in Primary Age-Related Tauopathy. medRxiv.
Publication Preprint 2023
Anatomic Landmarks Anti-Antibodies Antibodies Antigens CA1 Stratum Radiatum CA3 Stratum Lucidum Citrates Ethanol Formalin Goat Gyrus, Dentate IgG1 Immunoglobulins Lateral Geniculate Body Lipofuscin Mice, House Microscopy, Confocal Neoplasm Metastasis Neuropathologist Paraffin Pressure Rabbits Seahorses Synaptophysin Tissues
4
Adrenal Tissue Autofluorescence Quenching
Frozen mouse adrenal tissue sections were thawed for 30 min at RT and washed with PBS two times, 10 min each, to remove O.C.T. The tissue sections were then treated separately with tissue AF treatment methods (Table 1) at room temperature. Sudan Black B (SBB, Dia-m, Moscow, Russia) was prepared as 0.1% (W/V) in 70% ethanol, as described previously [27 (link)]. Sections were incubated with the SBB solution sealed airtight in the dark for 20 min, and then dipped briefly in 70% ethanol once before washing with PBS. A solution of 10 mM copper(II) sulfate (CuSO4) in 50 mM ammonia acetate, pH 5, was prepared and applied to sections for 90 min [27 (link)]. Ammonia/ethanol (NH3) was prepared as 0.25% (V/V) ammonia (PanReac AppliChem ITW Reagents, Barcelona, Spain) in 70% ethanol and applied to tissue sections for 1 h [29 (link)]. A fresh 0.05% (W/V) trypan blue (Paneko, Moscow, Russia) in PBS solution was prepared and applied to slides for 15 min [28 (link)]. The 20X TrueBlack solution (TrueBlackTM Lipofuscin Autofluorescence Quencher, Biotium, Fremont, CA, USA) was diluted to 1X in 70% ethanol and applied to tissue sections for 1 min. After each of these treatments, the slides were washed with PBS 3 times for 15 min each. TrueVIEW Reagent (TrueVIEWTM Autofluorescence Quenching Kit, Vector Laboratories, Burlingame, CA, USA) was prepared according to manufacturer instructions, applied immediately to sections for 3 min, and then washed once with PBS for 5 min. MaxBlockTM Autofluorescence Reducing Reagent Kit (MaxVision Biosciences, Bothell, WA, USA) was applied according to manufacturer instructions. Tissue sections were incubated with MaxBlockTM Autofluorescence Reducing Reagent (Reagent A) for 1 min, washed according to manufacturer instructions, incubated with Post-Detection Conditioner (Reagent B) for 5 min, and then washed according to manufacturer instructions. After each treatment, the slides were mounted onto coverslips with a polyvinyl alcohol mounting medium with DABCOTM antifading (Sigma-Aldrich, St. Louis, MO, USA). The slides treated with TrueVIEWTM Autofluorescence Quenching Kit were mounted with VECTASHIELD Vibrance Antifade Mounting Medium (Vector Laboratories, Burlingame, CA, USA) according to the manufacturer’s instructions. The slides were stored at 4 °C in the dark for subsequent examination.
Sakr N., Glazova O., Shevkova L., Onyanov N., Kaziakhmedova S., Shilova A., Vorontsova M.V, & Volchkov P. (2023). Characterizing and Quenching Autofluorescence in Fixed Mouse Adrenal Cortex Tissue. International Journal of Molecular Sciences, 24(4), 3432.
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Publication 2023
Acetate Aftercare Ammonia Cloning Vectors Copper Ethanol Frozen Sections Lipofuscin Mus Polyvinyl Alcohol Sudan Black B Sulfates, Inorganic Tissues Trypan Blue
5
Quantifying Intestinal Autofluorescence in Nematodes
Intestinal autofluorescence caused by lysosomal deposits of lipofuscin can accumulate over time in aging nematodes or nematodes exposed to specific toxicants, and the analytical method was performed as described previously [37 (link),38 (link)]. The exposed nematodes were anaesthetised with levamisole solution (60 µM), and then pipetted onto a slide containing a 2% agarose pad. Fluorescence intensity images of each group of nematodes intestinal autofluorescence were taken under a fluorescence microscope (UV-2A filter). The images were analysed by ImageJ V1.8.0 software and the results were expressed as mean fluorescence intensity values. A minimum of 30 nematodes were detected in each group, and the process was repeated three times.
Yang R., Ge P., Liu X., Chen W., Yan Z, & Chen M. (2023). Chemical Composition and Transgenerational Effects on Caenorhabditis elegans of Seasonal Fine Particulate Matter. Toxics, 11(2), 116.
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Publication 2023
Fluorescence Intestines Levamisole Lipofuscin Lysosomes Microscopy, Fluorescence Nematoda Sepharose

Top products related to «Lipofuscin»

1
Trueblack lipofuscin autofluorescence quencher by Biotium
Sourced in United States
TrueBlack Lipofuscin Autofluorescence Quencher is a reagent designed to reduce lipofuscin-based autofluorescence in biological samples. It functions by selectively quenching the fluorescent signal from lipofuscin without affecting the fluorescence of other cellular components.
2
Sudan black b by Merck Group
Sourced in United States, Germany
Sudan Black B is a laboratory reagent used for staining lipids and lipid-containing structures in biological samples. It is a lipophilic dye that binds to neutral lipids, providing a blue-black coloration. The dye is commonly used in histological and cytological procedures to visualize the presence and distribution of lipids in cells and tissues.
3
Trueblack by Biotium
Sourced in United States
TrueBlack is a dark background reagent designed for use in fluorescence microscopy and flow cytometry applications. It is formulated to effectively quench autofluorescence, improving the signal-to-noise ratio and enhancing the contrast of fluorescent signals.
4
4 6 diamidino 2 phenylindole (dapi) by Thermo Fisher Scientific
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DAPI is a fluorescent dye used in microscopy and flow cytometry to stain cell nuclei. It binds strongly to the minor groove of double-stranded DNA, emitting blue fluorescence when excited by ultraviolet light.
5
Alexa 488 by Thermo Fisher Scientific
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Alexa 488 is a fluorescent dye used in various biological applications. It is a brighly fluorescent, green-emitting dye with excitation and emission maxima at 495 and 519 nm, respectively. Alexa 488 can be conjugated to biomolecules such as proteins, antibodies, or nucleic acids to enable their detection and visualization.
6
Immuno fluore mounting medium by ICN Biomedicals
Sourced in United States
Immuno-Fluore mounting medium is a product designed to preserve and protect fluorescence signals in immunofluorescence applications. It is a clear, aqueous-based solution that helps maintain the integrity and brightness of fluorescent labels during microscopic analysis.
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Tcs sl confocal microscope by Leica
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The Leica TCS-SL confocal microscope is a high-performance, compact, and versatile laser scanning confocal imaging system designed for a wide range of applications. It features a fully automated optical system, advanced optics, and powerful imaging software to provide researchers with high-quality, high-resolution images.
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Vectashield mounting medium by Vector Laboratories
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Vectashield mounting medium is a proprietary aqueous-based formulation used for the preservation and visualization of fluorescent stained specimens. It is designed to maintain the brightness and stability of fluorescent dyes during microscopic examination.
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Bovine serum albumin (bsa) by Merck Group
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Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
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Lsm 700 by Zeiss
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The LSM 700 is a versatile laser scanning microscope designed for high-resolution imaging of samples. It provides precise control over the illumination and detection of fluorescent signals, enabling detailed analysis of biological specimens.

More about "Lipofuscin"

Lipofuscin is a pigmented substance that accumulates within various cell types, particularly post-mitotic cells like neurons and cardiomyocytes.
It is a byproduct of normal cellular metabolism and is considered a hallmark of aging.
Lipofuscin is composed of cross-linked proteins, lipids, and metals, and its buildup can impair cellular function, contributing to the development of age-related diseases.
Researchers studying lipofuscin can optimize their efforts with PubCompare.ai, an AI-powered tool that enhances reproducibility and accuracy by easily locating the best protocols from literature, preprints, and patents using AI-driven comparisons.
This streamlines the research process and helps researchers get the most accurate results.
Lipofuscin is also known as ceroid pigment or age pigment.
It can be visualized using various staining techniques, such as Sudan Black B, a lipophilic dye that binds to lipofuscin granules.
TrueBlack Lipofuscin Autofluorescence Quencher can also be used to reduce the autofluorescence of lipofuscin, which can interfere with other fluorescent labels like DAPI, Alexa 488, and Immuno-Fluore mounting medium.
Researchers can utilize confocal microscopy techniques, such as the TCS-SL confocal microscope or the LSM 700, to image and analyze lipofuscin accumulation within cells.
Bovine serum albumin (BSA) can be used as a blocking agent to reduce nonspecific binding during immunofluorescence experiments.
Vectashield mounting medium is another useful tool for preserving fluorescent signals and reducing photobleaching, which is important when studying the accumulation of lipofuscin over time.
By incorporating these techniques and tools, researchers can optimize their lipofuscin research and gain valuable insights into the role of this pigment in aging and age-related diseases.