Immunofluorescence staining was conducted as previously described. [21 (link)] Briefly, tissue sections (5 μm) were deparaffinized with xylene and rehydrated in a series of graded ethanols (100%, 75%, 50%). Microwave decloaking was performed in 10 mM sodium citrate (pH 6.0) and non-specific sites were blocked in TBSTw containing 1% Blocking Reagent (Roche Diagnostics, Indianapolis, IN), 5% normal goat or donkey sera, 1% bovine serum albumin fraction 5. Primary and secondary antibodies are listed in Table 1 . Immunofluorescent staining was imaged using a BZ-X710 digital microscope (Keyence, Itasca, IL, USA) fitted with at 20x (PlanFluor, NA 0.45) objective. Some tissue sections were imaged using an Eclipse E600 compound microscope (Nikon Instruments Inc., Melville, NY) fitted with a 20x dry objective (Plan Fluor NA = 0.75; Nikon, Melville, NY) and equipped with NIS elements imaging software (Nikon Instruments Inc.). Fluorescence was detected using DAPI (2-(4-amidinophenyl)-1H -indole-6-carboxamidine), FITC, Texas Red (Chroma Technology Corp, Bellows Fall, VT), and CY5 filter cubes (Nikon, Melville, NY).
Eclipse e600 compound microscope
Manufactured by Nikon
Sourced in United States
The Nikon Eclipse E600 is a compound microscope designed for laboratory use. It features a modular design and supports a range of optical configurations, including bright field, dark field, and phase contrast microscopy. The Eclipse E600 provides high-resolution imaging capabilities for a variety of sample types.
Automatically generated - may contain errors
Lab products found in correlation
Digital sight ds ri1, by Nikon (2 mentions)
Nis elements imaging software, by Nikon (1 mentions)
Blocking reagent, by Roche (1 mentions)
Bz x710 digital microscope, by Keyence (1 mentions)
Fluorescein isothiocyanate (fitc), by Chroma Technology (1 mentions)
Texas red, by Chroma Technology (1 mentions)
Nis elements br 3.2 imaging software, by Nikon (1 mentions)
Harris hematoxylin, by Thermo Fisher Scientific (1 mentions)
Vt1200, by Leica (1 mentions)
6 protocols using eclipse e600 compound microscope
1
Immunofluorescence Staining Protocol for Tissue Sections
2
Quantifying Proliferation and Apoptosis
3
Quantifying Prostate Cell Morphology
4
Insect Tissue Preparation and Imaging
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Samples were fixed in 2% PFA overnight and soaked in phenol for 6 days before paraffin embedding and cross-sectioning (as modified from Li et al. 2015) [23 (link)]. Prior to paraffin embedding, insects were re-fixed with formaldehyde then dehydrated in an ethanol gradient series. Samples were stored until sectioning. Transverse sections were taken using a Microm HM 325 rotary microtome (Walldorf, Germany) and slides hand-stained in Harris hematoxylin and eosin-phloxine. Slides were imaged using a Nikon Eclipse E600 compound microscope (Nikon Instruments, Melville, NewYork) and Nikon Digital Sight DS-Ri1 high-resolution microscope camera through Nikon NIS-Elements BR 3.2 imaging software.
5
Paraffin Sectioning of Beetle Heads
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Paraffin sectioning was completed according to the procedure in Li et al. (2015) (link). Heads were aseptically removed from the pronotum before sectioning. Once removed, beetle heads were fixed in 10% neutral buffered formalin (StatLab Medical Products) for 24 h and then soaked in phenol for 6 d at room temperature. Heads were treated with an automated tissue processor (Shandon Excelsior- Serial # EX01110212) to allow desiccation of tissues and infiltration of paraffin and subsequently embedded in paraffin blocks. We used a Microm HM 325 rotary microtome (Walldorf, Germany) to cut 10-μm transverse sections. Selected slides containing mycangia were confirmed by immediate observation, and were dried at 58°C in a lab oven (Boekel 107800) for 3 d, then stained with Harris-hematoxylin (Richard Allan Scientific # 7211) and eosin-phloxine (Surgipath # 080117), and examined and photographed using a Nikon Eclipse E600 compound microscope (Nikon Instruments, Melville, NY) with a Nikon Digital Sight DS-Ri1 high-resolution microscope camera to acquire brightfield images.
6
Fixation and Sectioning of Plant Pods
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Pods were fixed in 0.5% (w/v) paraformaldehyde in 100 mM sodium phosphate buffer (pH 7) for 45 min on ice and under vacuum. They were then washed three times in sodium phosphate buffer (pH 7) at room temperature. The fixed pods were embedded in 3% (w/v) agarose and sectioned to 40 µm using a Leica VT1200S vibrating microtome (Leica Microsystems, Germany). Pod sections were stained for 30 to 60 s at room temperature in 0.05% Toluidine Blue (pH 4.5) and then rinsed two times with distilled H2O. Sections were viewed on a Nikon Eclipse E600 compound microscope (Nikon Instruments Inc., Melville, USA).
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