Lung tumor bearing transgenic mice were randomized into 3 groups at least 5 animals for the following treatments: (1 (link)) no treatment (control); (2 (link)) 150 μg of pan-IR700 i.v. but no NIR light exposure (APC i.v. only); (3 (link))150 μg of pan-IR700 i.v. followed by NIR light at 50 J/cm2 on day 1 after intravenous injection of pan-IR700 (NIR-PIT). The mice were irradiated with NIR light from 2 directions (each 25 J/cm2) via the back and front using NIR laser light at 685 to 693 nm wavelength (BWF5-690-8-600-0.37; B&W TEK INC., Newark, DE, USA) (500 mW/cm2 × 50 s, 25 J/cm2) while the remainder of the body was shielded from light with aluminum foil. The power density of light in mW/cm2 was measured with an optical power meter (PM100, Thorlabs, Newton, NJ, USA). These therapies were performed every week for up to 3 weeks. Fluorescence images were not obtained because IR700 fluorescence accumulated within lung tumors was not detectable outside the body.
Bwf5 690 8 600 0
Manufactured by B&W Tek
Sourced in United States
The BWF5-690-8-600-0.37 is a fiber-coupled laser source from B&W Tek. It operates at a wavelength of 690 nm and has a maximum output power of 8 mW. The laser is coupled to a 600 μm core-diameter optical fiber with a numerical aperture of 0.37.
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10 protocols using bwf5 690 8 600 0
1
NIR-PIT for Lung Tumor Regression in Mice
2
NIR Laser-Activated Phototherapeutic Assay
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One hundred thousand cells were seeded into 24 well plates and incubated for 24 hr. Culture media was replaced with fresh culture media containing 10 µg/mL of tra-IR700 and the cells were incubated for 6 hr at 37°C. After washing with PBS, phenol red free culture medium was added. Then, cells were irradiated with NIR laser light at 685 to 695 nm wavelength (BWF5-690-8-600-0.37; B&W TEK INC., Newark, DE, USA). The actual power density of mW/cm2 was measured with an optical power meter (PM 100, Thorlabs, Newton, NJ, USA).
3
NIR Laser-Mediated Cell Irradiation
4
NIR Laser-Mediated Cell Irradiation
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One hundred thousand cells were seeded into 24 well plates or ten million cells were seeded onto a 10 cm dish and incubated for 24 hr. Medium was replaced with fresh culture medium containing 10 µg/mL of tra-IR700 which was incubated for 6 hr at 37°C. After washing with PBS, phenol red free culture medium was added. Then, cells were irradiated with a NIR laser, which emits light at 685 to 695 nm wavelength (BWF5-690-8-600-0.37; B&W TEK INC., Newark, DE, USA). The actual power density of mW/cm2 was measured with an optical power meter (PM 100, Thorlabs, Newton, NJ, USA).
5
Photodynamic Therapy Protocol with IR700 Conjugates
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In 24-well plates, 100 000
cells were seeded and incubated with each of the following cell line
and mAb-IR700 conjugate pairs (A431-luc-GFP, MDAMB468-luc-GFP: pan-IR700
or cet-IR700; 3T3/Her2-luc-GFP, Calu3-luc-GFP: tra-IR700) at 10 μg
mL–1 for 6 h at 37 °C. After the cells were
washed with PBS twice, PBS was added. Then, cells were irradiated
with either a red LED (L690-66-60; Marubeni America Co., Santa Clara,
CA) or laser (BWF5-690-8-600-0.37; B&W TEK INC., Newark, DE).
The power density was measured with an optical power meter (PM 100,
Thorlabs, Newton, NJ) to emit the same light dose (J cm–2) with either LED or laser, while the time of exposure was carefully
adjusted.
cells were seeded and incubated with each of the following cell line
and mAb-IR700 conjugate pairs (A431-luc-GFP, MDAMB468-luc-GFP: pan-IR700
or cet-IR700; 3T3/Her2-luc-GFP, Calu3-luc-GFP: tra-IR700) at 10 μg
mL–1 for 6 h at 37 °C. After the cells were
washed with PBS twice, PBS was added. Then, cells were irradiated
with either a red LED (L690-66-60; Marubeni America Co., Santa Clara,
CA) or laser (BWF5-690-8-600-0.37; B&W TEK INC., Newark, DE).
The power density was measured with an optical power meter (PM 100,
Thorlabs, Newton, NJ) to emit the same light dose (J cm–2) with either LED or laser, while the time of exposure was carefully
adjusted.
6
NIR Laser-Mediated Phototherapy Protocol
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One hundred thousand cells were seeded into 24-well plates, or ten million cells were seeded into a 10 cm dish and incubated for 24 hr. The medium was replaced with fresh culture medium containing 10 μg/mL of tra-IR700 that was incubated for 6 hr at 37°C. After washing with PBS, phenol red free culture medium was added. Then, cells were irradiated with a NIR laser, which emits light at a 685 to 695 nm wavelength (BWF5-690-8-600-0.37; B&W TEK INC., Newark, DE, USA). The output power density in mW/cm2 was measured with an optical power meter (PM 100, Thorlabs, Newton, NJ, USA).
7
Photodynamic Therapy Dose Comparison
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One hundred thousand cells were seeded into 24 well plates and incubated for 24 hr. Medium was replaced with fresh culture medium containing 10 μg/mL of pan-IR700 and incubated for 6 hr at 37°C. After washing with PBS, phenol red free culture medium was added. Then, cells were irradiated with either a red LED (L690-66-60; Marubeni America Co., Santa Clara, CA) or Laser (BWF5-690-8-600-0.37; B & W TEK INC., Newark, DE, USA). LED and Laser emit light at 670 to 710 nm and 685 to 695 nm wavelength, respectively. The power density was measured with an optical power meter (PM 100, Thorlabs, Newton, NJ, USA). To emit the same light dose (J/cm2) with either LED or Laser, the time of exposure was carefully adjusted.
8
In vitro NIR-PIT Through Bone
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In vitro NIR‐PIT trials were carried out through the bone sample with A431‐luc cells. Twenty‐four‐well plates were seeded with A431‐luc cells (1 × 105 per well) and, after 24 hours of incubation, pan‐IR700 was added to each well for a final concentration of 10 µg/mL. The cells were then incubated for an additional 3 hours at 37°C. The cells were covered by the bone sample and were irradiated with a laser (BWF5‐690‐8‐600‐0.37; B&W Tek), emitting light at 685‐695 nm wavelength at a power density of 373 mW/cm2 as measured with an optical power meter (PM100 and S310; Thorlabs). Near‐infrared light at the energy of 1, 2, 4, 8, 16, 32, 64, and 128 J/cm2 (3, 6, 11, 22, 43, 86, 172, and 344 seconds, respectively) was irradiated to each well one‐by‐one; the other wells were covered by aluminum foil.
9
NIR-PIT for Tumor Ablation
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Pan-IR700 (100 μg per 200 μl diluted with phosphate-buffered saline) was injected intravenously at 6 days after injection of the tumor cells. Fluorescence images were obtained with a Pearl Imager (LI-COR Biosciences) at 24h after injection of Pan-IR700 and immediately after NIR-PIT. The tumors on the right dorsum were NIR light exposed with NIR laser light at 685 to 693 nm wavelength (BWF5-690-8-600-0.37; B&W TEK INC., Newark, DE, USA) (500 mW/cm2 x 40 s, 20 J/cm2) while the left sided tumors and the remainder of the body were shielded from light with aluminum foil. The power density of light in mW/cm2 was measured with an optical power meter (PM100, Thorlabs, Newton, NJ, USA). This dose of 20 J/cm2 by NIR laser light was selected because it has been shown to reliably induce SUPR effects. 20J/cm2 induced by laser light is equivalent to 50 J/cm2 NIR light emitted by LED in this tumor model [9 (link)].
10
Near-infrared Photoimmunotherapy Efficacy
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