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In vivo rainbow Cytophone diagnostic platform. (a) The principle of two-color Cytophone; insets show a schematic of iRBC structure (top, left) and a photothermal (PT) image of an actual iRBC (bottom, right) with Hz peaks. (Adobe Fireworks CS5, www.adobe.com) (b). Time-color coding of two high-repetition-rate laser pulses with different wavelengths of λ1 and λ2—(top)—for spectral identification of iRBCs (middle) in the background of synchronized (e.g., from pigmented skin) and unsynchronized false signals (bottom). (c) Absorption spectra of melanin, nRBCs, Hz, indocyanine green (ICG), and magnetic beads14–21. (d) PA traces with positive and negative PA contrast against a blood background created by many nRBCs and circulating blood clots (CBCs).
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In vivo, Cytophone has demonstrated the capability for the early diagnosis of cancer, infection, and cardiovascular disorders through photoacoustic detection of circulating disease markers directly in the bloodstream with an unprecedented 1,000-fold improvement in sensitivity. Nevertheless, a Cytophone with higher specificity and portability is urg...
Citations
... For circulating tumor cell (CTC) detection in melanoma patients, we demonstrated a 1000fold sensitivity improvement over conventional diagnostics 38,40 . We previously demonstrated the successful application of PAFC for iRBC detection in vitro and in murine malaria models 37,41 . Here, we present the results of a cross-sectional and longitudinal diagnostic performance and safety study of Cytophone in adults presenting with and treated for uncomplicated malaria in Yaoundé, Cameroon. ...
... Testing of both malaria-uninfected individuals in endemic areas and the characterization of signals arising from free Hz or intraleukocytic Hz 51 in vivo in humans is underway and will allow further refinements in Cytophone's capabilities. Due to the differences in light absorption between plasma, RBCs, and leukocytes, we anticipate that the Cytophone will be able to distinguish between these potential sources of Hz 41 . The ability of Cytophone to quantify parasitemia will also be critical. ...
... Finally, further work is underway to assess the ability to both detect and distinguish all human malaria species. Based on our prior in vivo murine studies, we anticipate that this will be feasible due to differences based on the size and shape of Hz crystals, iRBC size (reticulocyte versus normocyte), possible tiny spectral Hz fingerprints, and circulating parasite life cycle stages, whose Hz contents is expected to differ between species 41,53 . Variations in Hz crystal size, shape, and distribution will also contribute to the Cytophone's ability to distinguish between ring-stage parasites, circulating gametocytes, and sequestered trophozoites. ...
Current malaria diagnostics are invasive, lack sensitivity, and rapid tests are plagued by deletions in target antigens. Here we introduce the Cytophone, an innovative photoacoustic flow cytometer platform with high-pulse-rate lasers and a focused ultrasound transducer array to noninvasively detect and identify malaria-infected red blood cells (iRBCs) using specific wave shapes, widths, and time delays generated from the absorbance of laser energy by hemozoin, a universal biomarker of malaria infection. In a population of Cameroonian adults with uncomplicated malaria, we assess our device for safety in a cross-sectional cohort (n = 10) and conduct a performance assessment in a longitudinal cohort (n = 20) followed for 30 ± 7 days after clearance of parasitemia. Longitudinal cytophone measurements are compared to point-of-care and molecular assays (n = 94). Cytophone is safe with 90% sensitivity, 69% specificity, and a receiver-operator-curve-area-under-the-curve (ROC-AUC) of 0.84, as compared to microscopy. ROC-AUCs of Cytophone, microscopy, and RDT compared to quantitative PCR are not statistically different from one another. The ability to noninvasively detect iRBCs in the bloodstream is a major advancement which offers the potential to rapidly identify both the large asymptomatic reservoir of infection, as well as diagnose symptomatic cases without the need for a blood sample.
... Currently, there are several common approaches to the construction of in vivo cytometry systems, including photoacoustic flow cytometry (PAFC) [8][9][10][11], ultrafast scanning photoacoustic system [10,12,13], fluorescence [14][15][16][17][18], fluorescence imaging [19][20][21][22][23], scattering [24][25][26][27], diffuse fluorescence [7,[28][29][30][31], stimulated Raman scattering [32] among other cytometry methods. ...
Photoacoustic flow cytometry is one of the most effective approaches to detect "alien" objects in the bloodstream, including circulating tumor cells, blood clots, parasites, and emboli. However, the possibility of detecting high-amplitude signals from these objects against the background of blood depends on the parameters of the laser pulse. So, the dependencies of photoacoustic signals amplitude and number on laser pulse energy (5-150 μJ), pulse length (1, 2, 5 ns), and pulse repetition rate (2, 5, 10 kHz) for the melanoma cells were investigated. First, the PA responses of a melanoma cell suspension in vitro was measured to directly assess the efficiency of converting laser light into an acoustic signal. After it the same dependence with the developed murine model based on constant rate melanoma cell injection into the animal blood flow was tested. Both in vivo and in vitro experiments show that signal generation efficiency increases with laser pulse energy above 15 μJ. Shorter pulses, especially 1 ns, provide more efficient signal generation as well as higher pulse rates. A higher pulse rate also provides more efficient signal generation, but also leads to overheating of the skin. The results show the limits where the photoacoustic flow cytometry system can be effectively used for detection of circulating tumor cells in undiluted blood both for in vitro experiments and for in vivo murine models. This article is protected by copyright. All rights reserved.
... Photoacoustic imaging can also be used to detect periodontitis and monitor gingival health [167] . For another, as a new diagnostic method, photoacoustic spectroscopy can be used for cancer therapy, renal failure, diabetes mellitus, autism, schizophrenia, and more [168][169][170] . ...
Breast cancer is the most common cancer, and triple-negative breast cancer (TNBC) has the highest metastasis and mortality rate among all breast cancer subtypes. Rujifang is a traditional Chinese medicine formula with many years of clinical application in breast cancer treatment. Here, we aim to investigate the effects of Rujifang on circulating tumor cell (CTC) dynamics and the tumor microenvironment in a ZsGreen/luciferase double-labeled TNBC orthotopic model. We report that the number of CTCs monitored by in vivo flow cytometry (IVFC) strongly correlates with disease progression. Rujifang treatment decreased the number of CTCs and suppressed the distant metastasis of TNBC. Moreover, immunofluorescence analysis revealed that Rujifang treatment could affect the tumor microenvironment by downregulating Kindlin-1, which has been reported to promote metastasis of TNBC. Our study provides evidence of the anti-metastatic effect of Rujifang against TNBC in an animal model using fluorescent cell lines. The results suggest the potential therapeutic value of Rujifang as an anti-metastatic drug, however, further clinical trials are needed to validate these findings in humans. This article is protected by copyright. All rights reserved.
Flow cytometry is a versatile tool with excellent capabilities to detect and measure multiple characteristics of a population of cells or particles. Notable advancements in in vivo photoacoustic flow cytometry, coherent Raman flow cytometry, microfluidic flow cytometry, etc. have been achieved in the last two decades, which endows flow cytometry with new functions and expands its applications in basic research and clinical practice. The advanced flow cytometry broadens the tools available to researchers to conduct research involving cancer detection, microbiology (COVID-19, HIV, bacteria, etc.), and nucleic acid analysis. This review presents an overall picture of advanced flow cytometers and provides not only a clear understanding of their mechanisms but also new insights into their practical applications. We identify the latest trends in this area and aim to raise awareness of advanced techniques of flow cytometry. We hope this review expands the applications of flow cytometry and accelerates its clinical translation. This article is protected by copyright. All rights reserved.
Integration of optical technologies in biomedical sciences permitted light manipulation at smaller time-length scales for specific detection and imaging of biological entities. Similarly, advances in consumer electronics and wireless telecommunications strengthened the development of affordable and portable point-of-care (POC) optical devices, circumventing the necessity of conventional clinical analyses by trained personnel. However, many of the POC optical technologies translated from bench to bedside require industrial support for their commercialization and dissemination to the population. This review aims to demonstrate the intriguing progress and challenges of emerging POC devices utilizing optics for clinical imaging (depth-resolved and perfusion imaging) and screening (infections, cancer, cardiac health, and hematolologic disorders) with a focus on research studies over the previous 3 years. Special attention is given to POC optical devices that can be utilized in resource-constrained environments. This article is protected by copyright. All rights reserved.
