Kl 1500 lcd

Animals underwent permanent focal ischemia by photothrombosis (PT) in the left hemisphere as previously described [12 (link), 35 (link)] or sham operation (same procedure as PT, but without illumination). During surgery, the body temperature of the animals was kept at 37 °C using a self-regulating heating pad. Briefly, isoflurane-anesthetized rats (2% in O₂ under spontaneous ventilation) received intravenous injections of the photosensitive dye Rose Bengal (10 mg/ml, Sigma, USA) in the tail vein. The skin above the skull was incised, and the brain was illuminated through the exposed skull with cold light (KL 1500 LCD, Schott) for 15 min at a stereotactically defined position (0.5 mm laterally and +4/−4 mm anterior/posterior to bregma), producing an approximate irradiation area of 8 × 4 mm². Incisions were sutured, and animals were allowed to awake from anesthesia while on a heating pad and returned to their home cages.

In all investigations, chick embryos at d16 were used because at this time point the blood volume approaches the maximum during the incubation period21 (link). After precooling, the shell access windows were enlarged without hurting the CAM to facilitate the injection of the CA. A Cold Light Source, KL 1500 LCD (SCHOTT, Mainz, Germany) was used for illumination. 30 G 1/2 needles (BD Microlance^TM 3, BD Drogheda, Ireland) and 0.01–1 ml injectors (B|BRAUN, Melsungen, Germany) were used for injection. CA was injected into a chorioallantoic capillary vein of medium size. After 5 minutes hemostasis by compression with cotton swab, about 200 μl OpSite* (Smith & Nephew, London, England) was applied to stop bleeding completely after injection. After ensuring that the bleeding was totally controlled, eggs were put back into the refrigerator.

All experiments were carried out with the approval of the Malmö-Lund animal review board (ethical permit number: M 25-12) and according to the ARRIVE guidelines. Animals were housed under reverse light conditions, with the testing performed during the dark period when the rats are active. The experiments were carried out on male Sprague Dawley rats (8 weeks, Charles River). The animals were anesthetized by isoflurane (approximately 2% in O₂ under spontaneous ventilation) and placed into a stereotaxic frame. Temperature was monitored during surgery using a rectal temperature probe and animals were kept at 37.0–37.5±0.2°C by means of a heating pad with feedback control (Table S1). A sagittal skin incision was made, subcutaneous connective tissue was removed and the skull was dried. Thereafter, the dye Rose Bengal (0.5 mL at 10 mg/mL) was injected in the tail vein. Two minutes after injection, the skull was illuminated with cold light (Schott, KL 1500 LCD) on an area of 8 to 4.5 mm for 15 min (from +4 to −4 mm antero-posterior and from 0.5 to 5 mm on the left from bregma). The tail and the scalp incisions were sutured and the rats transferred to their home cage. The functional deficit was assessed 2d after the onset of the stroke using the limb placement test and then regularly once a week.

We constructed a bespoke Y-maze with a short central arm (2×2.5 cm, length×width) and two choice arms (10×2.5 cm, length×width) separated by a 120 deg angle (Fig. 1). A single green LED with a narrow emission spectrum (555 nm; SSL-LX5093PGD, Lumex; Fig. S1A) was mounted 0.5 cm from the end of each choice arm. Males were allowed to explore the Y-maze for 5 min before the start of the experiment. Then, trials began with the male placed in the central arm so that both choice arms were visible. An LED was turned on in one choice arm at random; the intensity from the male starting location was 1 lx. Each trial lasted 5 min, after which the LED was turned off, the glow-worm returned to the starting position and the LED in the opposite arm turned on. Each experiment consisted of six sequential trials executed in a Y-maze unlit other than the green LED or illuminated with diffuse ‘warm’ white light (KL1500 LCD, Schott, Mainz, Germany; Fig. S1B) from 25 cm above with increasing intensity (25, 45, 90 and 145 lx) during the second to fifth trials. In some trials the white light was focused on the final 2.5 cm of each choice arm of the Y-maze, though there was no significant difference between the results of these trials and those with diffuse light (e.g. Fig. S2). Each set of six trials involved a different male glow-worm.

For all pre‐hatching stages, we used time‐lapse videography of embryos at room temperature with epi‐illumination from a fiber‐optic lamp (Schott KL 1500 LCD). Photos were taken every 5 min with a CCD (charge‐coupled device) camera (Nikon DS‐Fi1‐L2) connected to stereo microscope (Nikon SMZ1500). The images acquired had a minimum resolution of 300 dpi (dots per inch) and were stored in JPG format. During the recording, embryos were kept in glass embryo dishes (uncovered, 30 mm diameter × 12 mm deep) filled with embryo water. Because young embryos do not yet show spontaneous movements, it was not necessary to immobilize them with agarose embedding or anesthesia. The water level during the recording period was maintained by adding drops of egg water to the embryo dishes as necessary.

Male non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice (3–4 weeks old) were provided by HFK Bioscience Co. Ltd. (Beijing, China) and maintained under specific pathogen-free conditions. All animal experiments were performed in accordance with protocols evaluated and approved by the ethical committee of Tongji Medical College.
Inoculations with an intradermal injection of 1.2×10^⁷ Jeko-1 cells or 6×10^⁶ Raji cells were performed on the depilated right shoulders of NOD/SCID mice. PDT was conducted when tumors reached a diameter of 7 to 10 mm. A fiber optic bundle of a cold light (540 nm) source (KL 1500 LCD, SCHOTT, Germany), with a 4.5 mm aperture, was suspended 1 mm above the tumors to illuminate them for 15 min at an intensity of 3000 K; illumination started 15 min after the intravenous injection of nanoparticles. Thirty mice were then divided into six groups at random (n=5): normal saline, EGFP-EGF1-conjugated PEG-PLGA nanoparticle-loaded HMME (ENPs), PEG-PLGA nanoparticle-loaded HMME (NPs) plus PDT, catalase and HMME-encapsulated PEG-PLGA nanoparticles (CNPs) plus PDT, ENPs plus PDT, and CENPs plus PDT. Two mg kg⁻¹ of nanoparticles were injected into corresponding groups intravenously.

Living embryos were transferred into 2.5% methylcellulose (in E3-buffer) onto an object slide and image documentation was done with the bright-field stereomicroscope LEICA MZFLIII (Wetzlar, Germany) using the digital LEICA DFC310 FX camera, with the cold-light source KL 1500 LCD (Schott, Mainz, Germany). Video documentation for the beating hearts was performed using the LEICA DM IRB microscope with a magnification of 1.6×, a LEICA 506,059 objective, and a LEICA DFC360 FX camera (MCU II, Kappa GmbH, Germany). Images were processed with ImageJ 1.50i (National Institutes of Health, MD). Heart rate was calculated as heart beats per min. The “Zebrafs” program was used to analyze the percentage of ventricular and arterial fractional shortening.