Gutierrez-Juarez Gerardo, Gupta Sagar K, Al-Shaer Mays, Polo-Parada Luis, Dale Paul S, Papageorgio Chris, Viator John A
University of Guanajuato, Leon, Mexico.
Lasers Surg Med. 2010 Mar;42(3):274-81. doi: 10.1002/lsm.20894.
Circulating tumor cells have been shown to correlate positively with metastatic disease state in patients with advanced cancer. We have demonstrated the ability to detect melanoma cells in a flow system by generating and detecting photoacoustic waves in melanoma cells. This method is similar to flow cytometry, although using photoacoustics rather than fluorescence. Previously, we used piezoelectric films as our acoustic sensors. However, such films have indicated false-positive signals due to unwanted direct interactions between photons from the high laser fluence in the flow system and the film itself. We have adapted an optical detection scheme that obviates the need for piezoelectric films.
STUDY DESIGN/MATERIALS AND METHODS: Our photoacoustic system comprised a tunable laser system with an output of 410-710 nm with a pulse duration of 5 nanoseconds. The light was delivered by optical fiber to a glass microcuvette that contained saline buffer suspensions of melanoma and white blood cells. We used a continuous HeNe laser to provide a probe beam that reflected off of a glass and water interface in close proximity to the microcuvette. The beam was detected by a high-speed photodiode. When a photoacoustic wave was generated in the microcuvette, the wave propagated and changed the reflectance of the beam due to index of refraction change in the water. This perturbation was used to detect the presence of melanoma cells.
We determined a detection threshold of about one individual melanoma cell with no pyroelectric noise indicated in the signals.
The optical reflectance method provides sensitivity to detect small numbers of melanoma cells without created false-positive signals from pyroelectric interference, showing promise as a means to perform tests for circulating melanoma cells in blood samples.
循环肿瘤细胞已被证明与晚期癌症患者的转移疾病状态呈正相关。我们已经展示了通过在黑色素瘤细胞中产生和检测光声波,在流动系统中检测黑色素瘤细胞的能力。这种方法类似于流式细胞术,不过使用的是光声而不是荧光。此前,我们使用压电薄膜作为声学传感器。然而,由于流动系统中高激光能量密度的光子与薄膜本身之间不必要的直接相互作用,此类薄膜出现了假阳性信号。我们采用了一种光学检测方案,不再需要压电薄膜。
研究设计/材料与方法:我们的光声系统包括一个可调谐激光系统,输出波长为410 - 710纳米,脉冲持续时间为5纳秒。光通过光纤传输到一个玻璃微量比色皿,其中含有黑色素瘤细胞和白细胞的生理盐水缓冲液悬浮液。我们使用连续氦氖激光器提供探测光束,该光束从靠近微量比色皿的玻璃与水的界面反射。光束由高速光电二极管检测。当在微量比色皿中产生光声波时,波传播并由于水中折射率的变化而改变光束的反射率。这种扰动被用于检测黑色素瘤细胞的存在。
我们确定了大约单个黑色素瘤细胞的检测阈值,信号中未显示热释电噪声。
光学反射法能够灵敏地检测少量黑色素瘤细胞,且不会因热释电干扰产生假阳性信号,有望作为一种对血液样本中循环黑色素瘤细胞进行检测的手段。