We report a two-photon excitation fluorescence microscopy imager that uses a fiber-delivered fs supercontinuum (SC) generation source to achieve simultaneous dual-wavelength imaging. The SC generation is based on pumping a polarization-maintaining photonic crystal fiber with a fiber laser. The SC is then delivered to the imaging head through a broadband hollow-core fiber. Excitation wavelengths centered at 920 nm and 1180 nm were selected, which enabled simultaneous imaging of neurons transfected with GCaMP6 and administrated with muscimol-conjugated BODIPY. Imaging results of calcium dynamics of in vivo mouse brain and cultured primary neurons were collected to demonstrate the technology feasibility.
Intraoperative nerve recognition is critical to avoid accidental transection. Fluorescence-guided surgery can aid in nerve identification. However, detection of weak nerve-specific fluorescence signal is susceptible to the interference from high-background bright lights. We present a time-gated imager designed with ease-of-use and cost effectiveness in mind. Using this technology, we demonstrate successful rejection of room-light background signal to visualize the murine sciatic nerve.
Identification of positive margins during breast cancer conserving surgery is crucial for successful tumor resection. To address this clinical need, we developed a microscope that combines fluorescent microscopy, optical coherence tomography (OCT), and reflectance confocal microscopy (RCM) to identify the presence of positive tumor margins in surgical specimens. A cancer-targeting fluorescent agent is used to highlight potentially positive margins and guide combined RCM/OCT imaging. Combined RCM/OCT is used to accurately determine the extent of the positive margins, aided by custom image-processing techniques. This instrument has the potential to provide tumor margin guidance in the surgical suite, reducing reliance on lengthy histopathological inquiry and reducing the need for repeated surgery.
In the current era of personalized cancer therapy, various drugs need to be rapidly tested to determine their efficacy in killing cancer cells. In this way, the most effective drug can be administered to improve therapy effectiveness. However, since the biopsy specimens are heterogeneous, efficient testing requires the use only of the viable tumor part of the specimen. Therefore, rapid and automated evaluation of specimen heterogeneity is needed. In this paper presents an automated algorithm for tissue heterogeneity estimation based on OCT images. This algorithm use image texture features to differentiate between healthy and cancer tissue.
We report the development and the pre-clinical testing of a manual scanning OCT-based probe for core needle biopsy guidance. While stereotactic radiography, ultrasound, computed tomography, and magnetic resonance imaging are used to guide needle placement within a tumor, the optical probe provides the radiologist with the capability to examine tissue cellularity at the tip of the biopsy needle. The capability to investigate tissue cellularity prior to taking the biopsy could help reduce the number of non-diagnostic biopsies and increase the amount of viable tumor tissue within the biopsy core. This last aspect is very important in the new era of personalized cancer therapy, because greater tissue quantity is needed for various biomarker assays. This technology has been evaluated by us on a rabbit model of soft tissue cancer. Our results indicate the capability of this OCT-based probe for determining the in situ cellularity of the tissue at the tip of the biopsy needle.
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