Intensilight c hgfi mercury lamp

Samples were side-illuminated with a Nikon Intensilight C-HGFI mercury lamp. High angle scattered light is then collected by an M-Plan APO NUV 50X, NA 0.42 objective, which is transmitted via a FG600AEA Thorlabs fibre to a Princeton Instruments spectrometer.

All the measurements were performed under ambient environment and at room temperature conditions. Raman spectroscopy measurements were performed using a confocal scanning Raman microscope (WiTec Alpha300 system) in a backscattering geometry with 532 nm excitation (1 mW) and a ×100 objective (0.9 NA). Dark-field measurements were conducted in a backscattering geometry with illumination from a Nikon Intensilight C-HGFI mercury lamp, where the scattered light was collected using Nikon LU Plan ELWD ×100 (0.80 NA) objective and transmitted via a fibre to a single-photon-counting module (SPCM-AQRH) connected to a gated photon counter (SR400) or a Princeton Instruments spectrometer for microscopic and spectroscopic measurements, respectively. The transport measurements were conducted using a Keithley 2634B semiconductor parameter analyser unit with a two-point measurement for source and drain electrodes.

(i) H&E and PAS-AB. Mouse colons were placed intact in cassettes and fixed in 10% Carnoy’s fixative. Paraffin-embedded tissue sections (7 μm) were processed for hematoxylin and eosin (H&E) or periodic acid-Schiff/Alcian blue (PAS-AB) staining. H&E and PAS-AB sections were examined by bright-field and imaged on the Nikon Eclipse 90i (Nikon) microscope using a DS-Fi1-U2 camera (Nikon) with a differential interference contrast (DIC) objective.
(ii) Immunofluorescence.F. nucleatum localization was examined using a Fusobacterium-specific FISH probe (5′-CGCAATACAGAGTTGAGCCCTGC-3′), and total bacteria were examined using a universal bacterial FISH probe EUB338 (5′-GCTGCCTCCCGTAGGAGT-3′; Integrated DNA Technologies [IDT]) (110 (link)). Briefly, tissue sections were dehydrated and incubated with the Fusobacterium probe at 45°C in a dark humidifying chamber, hybridized for 45 min, and counterstained with MUC2 (1:200 dilution; Cloud-Clone Corp., PAA705Mu01) and Hoechst 33342 (Invitrogen, H3570). Immunostained slides were imaged on an Eclipse 90i (Nikon, Tokyo, Japan) with a 20× Plan Apo (NA 0.75) DIC objective, and the images were recorded using a CoolSNAP HQ2 camera (Photometrics) using a Nikon Intensilight C-HGFI mercury lamp.