Cells were plated onto glass cover slips in 24-well plates then fixed with formalin or methanol and permeabilized with Triton-X100 in PBS. Cover slips were blocked with fetal bovine serum in PBS or bovine serum albumin in PBST (0.1% Tween-20 in PBS) before incubating with primary and secondary antibodies in PBS. Cover slips were mounted onto microscope slides with Fluoroshield Mounting Medium containing DAPI (ab104139, Abcam). Primary antibodies used were anti-α-tubulin (1:2000, T3559, Sigma-Aldrich), anti-pericentrin (1:1500, ab4448, Abcam), anti-aurora kinase A (1:3000, ab13824, Abcam), anti-polo-like kinase 1 (1:100, sc-17783, Santa Cruz), and anti-kinesin family member 23 (1:500, HPA045208, Atlas antibodies). Secondary antibodies used were anti-mouse-488 (1:250, ab150117, abcam), anti-rabbit-594 (1:250, ab150080, abcam), and anti-rabbit-Cy3 (1:250, ab6939, abcam). Fluorescent images were acquired with an Axioplan 2 microscope (Carl Zeiss) and an AxioCam MRm Camera (Carl Zeiss), or an LSM 880 Airyscan microscope (Carl Zeiss).
Lsm 880 airyscan microscope
Manufactured by Zeiss
Sourced in Germany
The LSM 880 Airyscan microscope is a high-resolution confocal laser scanning microscope developed by Zeiss. It utilizes Airyscan technology to provide improved signal-to-noise ratio and higher resolution compared to traditional confocal microscopes. The core function of the LSM 880 Airyscan is to enable high-quality imaging of biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Electron Microscopy Sciences (4 mentions)
Softworx, by Cytiva (4 mentions)
Vectashield, by Vector Laboratories (3 mentions)
A1r microscope, by Nikon (3 mentions)
Deltavision omx 3d sim imaging system v4, by GE Healthcare (3 mentions)
Lsm 710, by Zeiss (3 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Tween 20, by Merck Group (2 mentions)
Spinning disk confocal microscope csu w1, by Zeiss (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Genejet transfection reagent, by SignaGen (2 mentions)
C apochromat 40x na 1.2, by Zeiss (2 mentions)
Zen black 2 3 sp1 software v14, by Zeiss (2 mentions)
No 1.5 borosilicate glass bottom, by Thermo Fisher Scientific (2 mentions)
Deltavision omx sr, by GE Healthcare (2 mentions)
Tirf sim microscope, by GE Healthcare (2 mentions)
Lsm 880, by Zeiss (2 mentions)
Zen 2.3 sp1 basic software, by Zeiss (2 mentions)
Zen 2.3 blue, by Zeiss (2 mentions)
Igg2α alexa fluor 647 secondary, by Thermo Fisher Scientific (2 mentions)
76 protocols using lsm 880 airyscan microscope
1
Immunofluorescence Microscopy of Cytoskeletal Proteins
2
Quantifying Invadopodia Length in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were grown on Oregon Green–coated coverslips for 4 h, fixed in room-temperature in 3% FA for 15 min and stained with anti-TKS5 and phalloidin/Alexa Fluor 647 as described above. Nuclei were stained in PBS/Hoechst 33342 (2 µg/ml) for 10 min, and the cells were mounted in ProLong Diamond. To measure the length of invadopodia, z-stacks of individual cells were acquired using a Zeiss LSM 880 Airyscan microscope in confocal mode, with a step size of 0.24 µm. The average invadopodia lengths from individual cells were calculated by defining the first and the last section from each z-stack that clearly showed a typical invadopodia staining of both TKS5 and actin and multiplying the number of sections with the step size.
3
High-resolution live-cell and fixed-oocyte imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Live-cell movies were acquired on a Leica SP5 confocal microscope using a 40x HCX PL AP 1.10 NA water immersion objective lens (Leica Microsystems), or a Zeiss LSM880 AiryScan microscope using a C-Apochromat 40 × 1.20 NA water immersion objective lens. For live-cell imaging experiments, at least 3–5 oocytes were recorded per session.
Fixed oocytes were imaged on a Leica SP8 microscope equipped with the HC PL APO 1.40 NA 100x oil immersion objective according to Nyquist criteria. For STED imaging, suitable Abberior STAR 580 and Abberior STAR RED or Abberior STAR 635P secondary antibodies or nanobodies were used (Abberior, NanoTag). Samples were imaged on a Leica SP8 STED microscope, with the HC PL APO CS2 1.40 NA 100x oil immersion objective and using the 775 nm depletion laser. Alternatively, we used an Abberior Instruments STEDYCON scan head mounted onto a Nikon Ti2 microscope equipped with a 100x CFI Plan Apochromat Lambda NA 1.45 oil immersion objective lens, or with an Abberior Instruments Expert Line STED microscope using an Olympus 100x UPLSAPO 100XS NA 1.4 oil immersion objective. At least five oocytes were recorded per sample.
Live and fixed oocyte images were processed and deconvolved using the Huygens software (Scientific Volume Imaging) with either confocal, AiryScan or STED settings as appropriate.
Fixed oocytes were imaged on a Leica SP8 microscope equipped with the HC PL APO 1.40 NA 100x oil immersion objective according to Nyquist criteria. For STED imaging, suitable Abberior STAR 580 and Abberior STAR RED or Abberior STAR 635P secondary antibodies or nanobodies were used (Abberior, NanoTag). Samples were imaged on a Leica SP8 STED microscope, with the HC PL APO CS2 1.40 NA 100x oil immersion objective and using the 775 nm depletion laser. Alternatively, we used an Abberior Instruments STEDYCON scan head mounted onto a Nikon Ti2 microscope equipped with a 100x CFI Plan Apochromat Lambda NA 1.45 oil immersion objective lens, or with an Abberior Instruments Expert Line STED microscope using an Olympus 100x UPLSAPO 100XS NA 1.4 oil immersion objective. At least five oocytes were recorded per sample.
Live and fixed oocyte images were processed and deconvolved using the Huygens software (Scientific Volume Imaging) with either confocal, AiryScan or STED settings as appropriate.
4
High-resolution Imaging of Actin Cytoskeleton
5
Immunofluorescence Imaging of Drosophila Embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embryos were collected on apple juice-agar plates and aged till the appropriate stage, dechorionated in 50% bleach, fixed in 1:1 heptane:4% formaldehyde in 1XPBS for 25 mins, devitellinized in a 1:1 mixture of methanol:heptane, and stored at −20°C in methanol. Embryos were rehydrated by washing in 1xPBS+0.2% Triton (PBT), blocked for ~1hr with 2% BSA in PBT, then incubated with the primary antibody at 4°C overnight. Following washes with PBT, embryos were incubated with the appropriate secondary antibody at room temperature for 2 hrs, stained in DAPI, and mounted onto a slide using VectaShield (Vector Laboratories) mounting medium. All primary antibodies were used at 1:250 dilution and secondaries at 1:1000. Imaging was performed on a Zeiss LSM880 Airy Scan microscope (Airy Fast mode) with 63X oil immersion objective at room temperature.
6
Immunofluorescence Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed in 4% paraformaldehyde (Electron Microscopy Sciences) for 10 min at room temperature; permeabilized in 100 mM tris-HCl (pH 7.4), 50 mM EDTA (pH 8.0), and 0.5% Triton X-100; and incubated with the corresponding primary antibodies overnight (see table S2). This was followed by incubation with corresponding fluorescently conjugated Alexa Fluor secondary antibodies for 2 hours at room temperature. Both primary and secondary antibodies were diluted in PBS containing 1% BSA (Sigma-Aldrich) and 0.1% Tween 20 (Sigma-Aldrich). Images were acquired using either a Nikon spinning disk confocal microscope (CSU-W1) or a Zeiss LSM880 Airyscan microscope. See table S2 for antibody information.
7
3D Bioprinting of GelMA-Nanoliposomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GelMA embedded, DiO-loaded nanoliposomes bioink (containing Irgacure 2959 photoinitiator, 0.1% in PBS pH = 7.4) was used to 3D bioprint disc-shaped constructs using a pneumatic extrusion bioprinter INKREDIBLE+ (CELLINK, Gothenburg, Sweden) equipped with a 23G nozzle, operating at pressures ranging from 60–70 kPa. GelMA-nanoliposomes were UV crosslinked (360–480 nm) for 40 s to generate nanocomposite hydrogels (Omnicure S-2000, 0.86 W/cm2). Confocal laser scanning microscopy imaging was performed in an LSM 880 Airyscan microscope (Carl Zeiss, Oberkochen, Germany) equipped with GaAsP/PMT detectors and a 20x/NA 0.8 Plan-Apochromat objective. Acquired data was post-processed in Zeiss ZEN v2.3 blue edition software.
8
Immunocytochemical Quantification of MUC5B
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fixed cells were washed once with PBS and permeabilized with PBS + 0.25% TritonX-100 (#1001541166, Sigma-Aldrich) for 20 min. The inserts were washed three times and blocked with PBS + 1% BSA (#1002119388, Sigma Aldrich) + 2% goat serum (#S-1000, Vector labs) at + 4 °C overnight, then incubated with PBS + 1% BSA + 2% goat serum + 5 µg/ml anti-MUC5B (#ab77995, Abcam) for 2 h. The cells were washed four times for 5 min and stained with PBS + goat anti-mouse IgG Alexa Fluor568 (#A21124, Invitrogen, 1:400) + Alexa Fluor 488 Phalloidin (#A12379, Invitrogen, 1:50) for 1 h, protected from light. The inserts were washed two times for 5 min and counterstained with PBS + Hoechst 33342 (#62249, Thermo Fisher Scientific, 1 µg/ml) for 10 min. After four 5-min washes, the membranes were removed from the culture inserts and mounted using Vectashield mounting media (#H-1000, Vector labs). Three representative images/membrane were captured using an LSM 880 Airyscan microscope (Zeiss) and the percentage of MUC5B/total image area was quantified using the Visiopharm Integrator System (Visiopharm A/S). All immunocytochemistry data are presented as mean of three independent experiments using cell-free conditioned media from three different AM donors and triplicate technical replicates.
9
Visualizing E3 Ligase Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells plated on a coverslip were transfected the next day with plasmids expressing 3xFLAG-E3 ligases for 24 h. To visualize the expression of the E3 ligases, cells were washed with PBS, fixed with 4% paraformaldehyde (PFA) in 1xPBS, permeabilized with 0.2% Triton in 1xPBS and blocked in 8% Bovine Serum Albumin (BSA) in 1xPBS for 1 h. Then, cells were incubated with primary anti-FLAG antibody (1/2000, Sigma, Cat#F1804) in 5% BSA-1xPBS for 1 h at room temperature (RT, 25°C). After 3 washes with 1xPBS, secondary anti-mouse antibody coupled to Alexa Fluor 488 diluted in 5% BSA-1xPBS (1/1000, ThermoFischer Scientific, Cat#A11001) was added for 2 h at RT. Nucleus were marked with 4′,6-diamidino-2-phenylindole (DAPI) and coverslips were mounted using a fluorescence mounting medium (Dako, Cat#S3023). Images were obtained using a confocal LSM880 Airyscan microscope (Zeiss) and analyzed with Fiji software.
10
Immunohistochemical Profiling of P2Y Receptors in the Organ of Corti
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Inner ears were dissected out and perfused with 4% paraformaldehyde solution for 20 min at room temperature. The fixed samples were then micro‐dissected to retrieve the apical turn of the organ of Corti. These were then blocked in 5% horse serum for an hour at room temperature before being incubated at 35°C overnight in primary antibody solution. The primary antibodies used included anti‐P2Y1 (Alomone Labs, cat. no. APR‐009), anti‐P2Y2 (Alomone Labs, cat. no. APR‐010) and anti‐P2Y4 (Alomone Labs, cat. no. APR‐006) all at a concentration of 1:800. The following day, samples were washed in phosphate buffer saline (PBS) and incubated for an hour at 35°C in secondary antibody solution. This solution contained goat anti‐rabbit IgG 490 (Alexa fluor 488, A11034, Thermo Fisher) and Texas Red‐X Phalloidin for staining of F‐actin (1:1000, T7471, Thermo Fisher). The samples were then washed in PBS and mounted in VECTASHIELD (H‐1000). Images of the cochleae were taken with a Zeiss LSM 880 Airyscan microscope. The z‐stack images were taken with 0.5 μm incremental steps. Fiji ImageJ software was used to process the images and generate maximum intensity projections (https://imagej.net/Fiji ).
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!