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Feedback-based wavefront shaping is a promising and versatile technique for enhancing the contrast of a target signal for both coherent and incoherent light through a highly scattering medium. However, this technique can fail for a dynamic sample with a short correlation time. So far, most proposed methods for high-speed wavefront shaping can only directly enhance the intensity of coherent light but not incoherent light. Here we try to fill this gap to directly enhance incoherent light with high speed, such as fluorescence, which is essential in extending wavefront shaping to biomedical applications. For this purpose, we develop a technique based on a single acousto-optic deflector (AOD) with field-programmable gate array (FPGA) acceleration for spatiotemporal focusing within milliseconds. With the digital time gating of the feedback signal, spatiotemporal focusing of laser light with high contrast can be formed behind dynamic scattering media in milliseconds resulting in fluorescence enhancement. Furthermore, FPGA-based wavefront shaping is shown to effectively enhance fluorescence directly behind dynamic samples with short correlation times.
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This publisher's note contains corrections to Opt. Lett.47, 5192 (2022)10.1364/OL.467435.
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Feedback-based wavefront shaping is a promising and versatile technique for enhancing the contrast of a target signal through highly scattering media. However, this technique can fail for low optical signals such as fluorescence and Raman signals or in a reflection setup because the trend in weak feedback signals can be easily overwhelmed by noise. To address this challenge, we develop a technique based on a single acousto-optic deflector (AOD) to create a signal with a selected beat frequency from optical signals that can serve as feedback, in which the phase distribution of various radio frequency components of the driving signal for the AOD is optimized for wavefront shaping. By shifting incident light frequency with the AOD, the feedback signal at a selected beat frequency can be measured with a high signal-to-noise ratio (SNR) by a lock-in amplifier, thus enabling the enhancement of weak target signals through highly scattering media. It is found that the method of lock-in beat frequency detection can significantly improve fluorescence imaging and Raman spectral measurements in a reflection setup, and thus could be potentially used for in vivo measurements.
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Imagen Óptica , Retroalimentación , Imagen Óptica/métodos , Relación Señal-RuidoRESUMEN
Lipid droplets have gained strong interest in recent years to comprehend how they function and coordinate with other parts of the cell. However, it remains challenging to study the regulation of lipid droplets in live preadipocytes using conventional microscopic techniques. In this paper, we study the effects of fatty acid stimulation and cell starvation on lipid droplets using optical diffraction tomography and Raman spectroscopy by measuring size, refractive index, volume, dry mass and degree of unsaturation. The increase of fatty acids causes an increase in the number and dry mass of lipid droplets. During starvation, the number of lipid droplets increases drastically, which are released to mitochondria to release energy. Studying lipid droplets under different chemical stimulations could help us understand the regulation of lipid droplets for metabolic disorders, such as obesity and diabetes.
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Adipocitos/metabolismo , Gotas Lipídicas/metabolismo , Espectrometría Raman/métodos , Tomografía Óptica/métodos , Células 3T3-L1 , Animales , Calibración , Holografía , Ratones , Tamaño de la Partícula , Imagen de Lapso de TiempoRESUMEN
We report a joint system with both confocal Raman spectroscopy (CRS) and optical coherence tomography (OCT) modules capable of quickly addressing the region of interest in a tissue for targeted Raman measurements from OCT. By using an electrically tunable lens in the Raman module, the focus of the module can be adjusted to address any specific depth indicated in an OCT image in a few milliseconds. We demonstrate the performance of the joint system in the depth dependent measurements of an ex vivo swine tissue and in vivo human skin. This system can be useful in measuring samples embedded with small targets, for example, to identify tumors in skin in vivo and assessment of tumor margins, in which OCT can be used to perform initial real-time screening with high throughput based on morphological features to identify suspicious targets then CRS is guided to address the targets in real time and fully characterize their biochemical fingerprints for confirmation.
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To monitor blood glucose levels reliably, diabetic patients usually have to undergo frequent fingerstick tests to draw out fresh blood, which is painful and inconvenient with the potential risk of cross contamination especially when the lancet is reused or not properly sterilized. This work reports a novel surface-enhanced Raman spectroscopy (SERS) sensor for the in situ intradermal detection of glucose based on a low-cost poly(methyl methacrylate) microneedle (PMMA MN) array. After incorporating 1-decanethiol (1-DT) onto the silver-coated array surface, the sensor was calibrated in the range of 0-20 mM in skin phantoms then tested for the in vivo quantification of glucose in a mouse model of streptozocin (STZ)-induced type I diabetes. The results showed that the functional poly(methyl methacrylate) microneedle (F-PMMA MN) array was able to directly measure glucose in the interstitial fluid (ISF) in a few minutes and retain its structural integrity without swelling. The Clarke error grid analysis of measured data indicated that 93% of the data points lie in zones A and B. Moreover, the MN array exhibited minimal invasiveness to the skin as the skin recovered well without any noticeable adverse reaction in 10 min after measurements. With further improvement and proper validation, this polymeric MN array-based SERS biosensor has the potential to be used in painless glucose monitoring of diabetic patients in the future.
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Técnicas Biosensibles , Espectrometría Raman , Animales , Glucemia , Automonitorización de la Glucosa Sanguínea , Glucosa , Humanos , RatonesRESUMEN
The progression of epithelial precancers into cancer is accompanied by changes of tissue and cellular structures in the epithelium. Correlations between the structural changes and scattering coefficients of esophageal epithelia were investigated using quantitative phase images and the scattering-phase theorem. An ex vivo study of 14 patients demonstrated that the average scattering coefficient of precancerous epithelia was 37.8% higher than that of normal epithelia from the same patient. The scattering coefficients were highly correlated with morphological features including the cell density and the nuclear-to-cytoplasmic ratio. A high interpatient variability in scattering coefficients was observed and suggests identifying precancerous lesions based on the relative change in scattering coefficients.