Tracing the formation and degradation of fatty-acid-rich mitochondria using label-free chemical imaging (Conference Presentation)

2020 ◽  
Author(s):  
Chi Zhang ◽  
Stephen A. Boppart
Keyword(s):  
Fatty Acid ◽  
Chemical Imaging ◽  
Label Free

Tracking the Formation and Degradation of Fatty-Acid-Accumulated Mitochondria Using Label-Free Chemical Imaging

2020 ◽  
Author(s):  
Chi Zhang ◽  
Stephen Boppart
Keyword(s):  
Fatty Acid ◽  
Structural Features ◽  
Chemical Imaging ◽  
Live Cells ◽  
Label Free ◽  
Dynamic Changes ◽  
Regulatory Processes ◽  
Transport Chain ◽  
Anti Stokes ◽  
First Time

Abstract The mitochondrion is one of the key organelles for maintaining cellular homeostasis. External environmental stimuli and internal regulatory processes alter the metabolism and functions of mitochondria. To understand these activities of mitochondria, it is critical to probe the key metabolic molecules inside these organelles. In this study, we used label-free chemical imaging modalities including coherent anti-Stokes Raman scattering and multiphoton-excited autofluorescence to study the mitochondrial activities in living cancer cells. We found that hypothermia exposure tends to induce fatty-acid (FA) accumulation in some mitochondria of MIAPaCa-2 cells. Autofluorescence images show that the FA-accumulated mitochondria also have abnormal NADH and FAD metabolism, likely induced by the dysfunction of the electron transport chain. We also found that when the cells were re-warmed to physiological temperature after a period of hypothermia, the FA-accumulated mitochondria changed their structural features, likely caused by the mitophagy process. To the best of our knowledge, this is the first time that FA accumulation in mitochondria was observed in live cells. Our research also demonstrates that multimodal label-free chemical imaging is an attractive tool to discover abnormal functions of mitochondria at the single-organelle level and can be used to quantify the dynamic changes of this organelle under perturbative conditions.

scholarly journals Tracking the formation and degradation of fatty-acid-accumulated mitochondria using label-free chemical imaging

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chi Zhang ◽  
Stephen A. Boppart
Keyword(s):  
Fatty Acid ◽  
Structural Features ◽  
Chemical Imaging ◽  
Live Cells ◽  
Label Free ◽  
Regulatory Processes ◽  
Transport Chain ◽  
Abnormal Metabolism ◽  
Anti Stokes ◽  
First Time

AbstractThe mitochondrion is one of the key organelles for maintaining cellular homeostasis. External environmental stimuli and internal regulatory processes may alter the metabolism and functions of mitochondria. To understand these activities of mitochondria, it is critical to probe the key metabolic molecules inside these organelles. In this study, we used label-free chemical imaging modalities including coherent anti-Stokes Raman scattering and multiphoton-excited fluorescence to investigate the mitochondrial activities in living cancer cells. We found that hypothermia exposure tends to induce fatty-acid (FA) accumulation in some mitochondria of MIAPaCa-2 cells. Autofluorescence images show that the FA-accumulated mitochondria also have abnormal metabolism of nicotinamide adenine dinucleotide hydrogen, likely induced by the dysfunction of the electron transport chain. We also found that when the cells were re-warmed to physiological temperature after a period of hypothermia, the FA-accumulated mitochondria changed their structural features. To the best of our knowledge, this is the first time that FA accumulation in mitochondria was observed in live cells. Our research also demonstrates that multimodal label-free chemical imaging is an attractive tool to discover abnormal functions of mitochondria at the single-organelle level and can be used to quantify the dynamic changes of these organelles under perturbative conditions.

scholarly journals Viewing Cancer in a New Light: Multimodal Label-Free Specific Chemical Imaging to Distinguish Healthy Cell Tissue from Invasive Carcinoma

2017 ◽  
Vol 112 (3) ◽  
pp. 187a
Author(s):  
Bjarne Thorsted ◽  
Stine R. Larsen ◽  
Christian Godballe ◽  
Jonathan R. Brewer
Keyword(s):  
Invasive Carcinoma ◽  
Chemical Imaging ◽  
Label Free ◽  
Specific Chemical ◽  
Healthy Cell ◽  
Cell Tissue

scholarly journals Whole organ cross-section chemical imaging using label-free mega-mosaic FTIR microscopy

The Analyst ◽  
2013 ◽  
Vol 138 (23) ◽  
pp. 7066 ◽  
Author(s):  
Paul Bassan ◽  
Ashwin Sachdeva ◽  
Jonathan H. Shanks ◽  
Mick D. Brown ◽  
Noel W. Clarke ◽  
...  
Keyword(s):  
Cross Section ◽  
Chemical Imaging ◽  
Label Free ◽  
Ftir Microscopy

Monitoring the effects of chemical stimuli on live cells with metasurface-enhanced infrared reflection spectroscopy

2021 ◽  
Author(s):  
Steven H. Huang ◽  
Jiaruo Li ◽  
Zhiyuan Fan ◽  
Robert Delgado ◽  
Gennady Shvets
Keyword(s):  
Intracellular Signaling ◽  
Strong Field ◽  
Detection System ◽  
Field Enhancement ◽  
Chemical Imaging ◽  
Live Cells ◽  
Label Free ◽  
Reflection Spectroscopy ◽  
Infrared Reflection ◽  
Infrared Reflection Spectroscopy

Infrared spectroscopy has found wide applications in the analysis of biological materials. A more recent development is the use of engineered nanostructures, or plasmonic metasurfaces, as substrates for metasurface-enhanced infrared reflection spectroscopy (MEIRS). Here, we demonstrate that strong field enhancement from plasmonic metasurfaces enables the use of MEIRS as a highly informative analytic technique for real-time monitoring of cells. By exposing live cells cultured on a plasmonic metasurface to chemical compounds, we show that MEIRS can be used as a label-free phenotypic assay for detecting multiple cellular responses to external stimuli: changes in cell morphology, adhesion, lipid composition of the cellular membrane, as well as intracellular signaling. Using a focal plane array detection system, we show that MEIRS also enables spectro-chemical imaging at the single-cell level. The described metasurface-based all-optical sensor opens the way to a scalable, high-throughput spectroscopic assay for live cells.

scholarly journals Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering

2019 ◽  
Vol 116 (32) ◽  
pp. 15842-15848 ◽  
Author(s):  
Yuta Suzuki ◽  
Koya Kobayashi ◽  
Yoshifumi Wakisaka ◽  
Dinghuan Deng ◽  
Shunji Tanaka ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Raman Scattering ◽  
Cancer Biology ◽  
High Speed ◽  
Stimulated Raman Scattering ◽  
Chemical Imaging ◽  
Fluorescent Labeling ◽  
Label Free ◽  
Imaging Flow Cytometry ◽  
Stimulated Raman

Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast-flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to ∼140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood.

scholarly journals Chemical imaging of live fibroblasts by SERS effective nanofilm

2014 ◽  
Vol 16 (44) ◽  
pp. 24621-24634 ◽  
Author(s):  
D. Radziuk ◽  
R. Schuetz ◽  
A. Masic ◽  
H. Moehwald
Keyword(s):  
Real Time ◽  
Chemical Imaging ◽  
Raman Imaging ◽  
Label Free

Label-free Raman imaging of live single NIH3T3 fibroblast produced by SERS effective SiO2@Ag–PAA nanoshells in real time.

scholarly journals Saliva-Omics in Plasma Cell Disorders — Proof of Concept and Potential As a Non-Invasive Tool for Monitoring Disease Burden and MRD Status

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 3177-3177
Author(s):  
Ciara Tierney ◽  
Despina Bazou ◽  
Giao Le ◽  
Paul Dowling ◽  
Peter O'Gorman
Keyword(s):  
Mass Spectrometry ◽  
Fatty Acid ◽  
Malignant Transformation ◽  
Fatty Acid Binding Protein ◽  
Proof Of Concept ◽  
Newly Diagnosed ◽  
Label Free ◽  
Fatty Acid Binding ◽  
Non Invasive ◽  
Acid Binding Protein

Abstract Salivaomics has exciting potential for the diagnosis and monitoring of malignancy, evidence of which has been reported in oral cancer (Sahibzada et al., 2017), head and neck malignancies (Citrin et al., 2012) and ovarian cancer (Chen et al., 1990). It has been observed that approximately 40% of cancer, stroke and cardiovascular disease biomarkers are present in whole saliva (Loo et al., 2010). Salivaomics has become an area of great interest in disease diagnosis over the last number of years, following the footsteps of the other "omics" based diagnostic tools. Saliva has been referred to as "the mirror of the body" as it gives an insight into the internal pathological state (Lee and Wong, 2009). As saliva is considered a fast, inexpensive and non-invasive method of sample collection, the future of diagnosis, early detection, monitoring and prediction of progression of disease has been thought to lie here. Monoclonal gammopathy of undetermined significance (MGUS) is characterised as a premalignant precursor tumours of MM. It has been seen that the majority of MM cases develop from MGUS (Weiss et al., 2009), leading to the need for biomarkers to monitor disease progression and explore the mechanism of malignant transformation. Serum and saliva samples were collected from 18 newly diagnosed MM patients and 8 MGUS patients, peptides were purified using the filter aided sample preparation (FASP) method and samples were prepared for label-free liquid chromatography mass spectrometry (LC-MS/MS) using an LTQ Orbitrap XL mass spectrometrer (Thermo Fisher Scientific). Proteins were analysed using the MaxQuant and Perseus software for mass-spectrometry (MS)-based proteomics data analysis, UniProtKB-Swiss Prot database and KEGG Pathway database. The abundance of proteins in saliva from MGUS compared to newly diagnosed MM was analysed using label-free mass spectrometry. A panel of 6 significant proteins was identified. Fatty Acid Binding Protein 5 (FABP5) was detected in elevated levels in saliva from MM patients compared to MGUS. The increased expression was verified using western blotting. Fatty Acid Binding Protein 5 (FABP5) is known to promote cell proliferation, survival and migration (Wang et al., 2006). FABP5 has been observed to aid in the proliferation of cancer and has been seen to be overexpressed in multiple cancer types such as breast (Levi et al., 2013), prostate (Morgan et al., 2008) and HCC (Ohata et al., 2017). FABP5 has seen to link closely with poor outcome and unfavourable clinical parameters in MM (Waheed et al., 2013). Additionally, analysis was performed on serial MM patient saliva samples during treatment. A panel of significant proteins was identified when comparing the saliva proteome during treatment the abundance of many of the detected proteins mirrored response to treatment (Table 1). For example, Transglutaminase 3 (TGM3) was observed as being decreased in abundance in saliva from patients in remission. TGM3 is expressed in brain, small intestine, mucosa and skin and has been shown to be required for the cross-linking of the structural protein trichohyalin and the keratin intermediate filaments to form a rigid structure within the inner root sheath cells (Hitomi, 2005). Overexpression of TGM3 has been observed to induce oesophageal cancer (EC) cell invasion, migration and proliferation. It has also been noted that TGM3 promotes apoptosis in EC in vitro (Li et al., 2016). TGM3 has been predicted to be a potential biomarker for disease progression in MM. This study provides proof of concept that a range of biologically significant proteins of interest can be reliably detected in the saliva of MM and MGUS patients. The observation of differential expression of FABP5 between MGUS and MM identified these as candidate proteins relevant to malignant transformation of MGUS to symptomatic MM. The demonstration of decreased abundance of FABP5 after achieving remission indicates a correlation with tumour burden. This opens the opportunity to explore candidate salivary biomarkers for use in the clinic for disease monitoring and Minimal Residual Disease (MRD) assessment. Disclosures No relevant conflicts of interest to declare.

scholarly journals High resolution mass spectrometry imaging of plant tissues: towards a plant metabolite atlas

The Analyst ◽  
2015 ◽  
Vol 140 (22) ◽  
pp. 7696-7709 ◽  
Author(s):  
Dhaka Ram Bhandari ◽  
Qing Wang ◽  
Wolfgang Friedt ◽  
Bernhard Spengler ◽  
Sven Gottwald ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectrometry Imaging ◽  
High Spatial Resolution ◽  
High Resolution Mass Spectrometry ◽  
Chemical Imaging ◽  
Plant Tissues ◽  
Label Free ◽  
Wide Range ◽  
Plant Metabolite ◽  
Resolution Mass

Label-free chemical imaging of a wide range of metabolites in all major plant organs acquired at high spatial resolution.

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