Regulation of lipid droplets in live preadipocytes using optical diffraction tomography and Raman spectroscopy

Hsieh, Chao-Mao; Liu, Patricia Yang; Chin, Lip Ket; Zhang, Jing Bo; Wang, Kuan; Sung, Kung-Bin; Ser, Wee; Bourouina, Tarik; Leprince‐Wang, Yamin; Liu, Ai Qun

doi:10.1364/oe.27.022994

Cited by 14 publications

(8 citation statements)

References 27 publications

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“…Using the developed system and fluorescence microscopy in combination, we performed experiments for cell biology studies. Nucleoli and lipid droplets were recognized as having greater OPL in QPI images (Figures 6–7, S2 and S3), consistent with previous reports (Hsieh et al, 2019; Imai et al, 2017; Liu et al., 2016; Shribak et al., 2017; Zhang et al., 2017). Nucleolus is known as a classical example of liquid–liquid phase separation (LLPS) (Brangwynne et al., 2011), and our system might be applicable to studies in this field.…”

Section: Discussionsupporting

confidence: 90%

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Single‐shot quantitative phase imaging as an extension of differential interference contrast microscopy

et al. 2021

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Various studies have been conducted to obtain quantitative phase information based on differential interference contrast (DIC) microscopy. As one such attempt, we propose in this study a single‐shot quantitative phase imaging (QPI) method by combining two developments. First, an add‐on optical system to a commercialized DIC microscope was developed to perform quantitative phase gradient imaging (QPGI) with single image acquisition using a polarization camera. Second, an algorithm was formulated to reconstitute QPI from the obtained QPGI by reducing linear artifacts, which arise in simply integrated QPGI images. To demonstrate the applicability of the developed system in cell biology, the system was used to measure various cell lines and compared with fluorescence microscopy images of the same field of view. Consistent with previous studies, nucleoli and lipid droplets can be imaged by the system with greater optical path lengths (OPL). The results also implied that combining fluorescence microscopy and the developed system might be more informative for cell biology research than using these methods individually. Exploiting the single‐shot performance of the developed system, time‐lapse imaging was also conducted to visualize the dynamics of intracellular granules in monocyte‐/macrophage‐like cells. Our proposed approach may accelerate the implementation of QPI in standard biomedical laboratories.

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Section: Discussionsupporting

confidence: 90%

“…Second, we focused on lipid droplets, because they are known to have higher RI in cytoplasm (Hsieh et al., 2019). HepG2 cells were double stained with Hoechst and LipiDye Ⅱ reagents to visualize specifically nuclei and lipid droplets, respectively.…”

Section: Resultsmentioning

confidence: 99%

Single‐shot quantitative phase imaging as an extension of differential interference contrast microscopy

et al. 2021

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“…In fact, adipogenesis is a mechanosensitive process, and increasing evidence links lipid production in adipocytes to a particular mechanical environment [ 46 ]. This circumstance, together with the differential effect of saturated and unsaturated FA on adipocyte maturation [ 47 ], allows us to speculate that differences between SOLF and UOLF mice regarding Lep gene expression previously reported by us [ 12 , 29 ] may be linked to the biophysical properties of the tissue [ 45 , 48 , 49 ].…”

Section: Discussionmentioning

confidence: 92%

Effect of Lauric vs. Oleic Acid-Enriched Diets on Leptin Autoparacrine Signalling in Male Mice

et al. 2022

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High-fat diets enriched with lauric acid (SOLF) do not enhance leptin production despite expanding white adipose tissue (WAT). Our study aimed at identifying the influence of SOLF vs. oleic acid-enriched diets (UOLF) on the autoparacrine effect of leptin and was carried out on eight-week-old mice consuming control chow, UOLF or SOLF. Phosphorylation of kinases integral to leptin receptor (LepR) signalling pathways (705Tyr-STAT3, 473Ser-Akt, 172Thr-AMPK), adipocyte-size distribution, fatty acid content, and gene expression were analyzed in WAT. SOLF enhanced basal levels of phosphorylated proteins but reduced the ability of leptin to enhance kinase phosphorylation. In contrast, UOLF failed to increase basal levels of phosphorylated proteins and did not modify the effect of leptin. Both SOLF and UOLF similarly affected adipocyte-size distribution, and the expression of genes related with adipogenesis and inflammation. WAT composition was different between groups, with SOLF samples mostly containing palmitic, myristic and lauric acids (>48% w/w) and UOLF WAT containing more than 80% (w/w) of oleic acid. In conclusion, SOLF appears to be more detrimental than UOLF to the autoparacrine leptin actions, which may have an impact on WAT inflammation. The effect of SOLF and UOLF on WAT composition may affect WAT biophysical properties, which are able to condition LepR signaling.

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“…Recently TPM has been demonstrated to be a powerful tool for the stain-free quantification of 3D features related to the cell biophysical properties. In particular, TPM has been already applied for LDs analysis (28)(29)(30)(31)(32)(33). However, so far TPM has been realized only in static environment for studying adhered cells, thus not overcoming the lowthroughput issue also typical of 3D confocal microscopy.…”

Section: Discussionmentioning

confidence: 99%

“…The first demonstration in visualizing and measuring LDs in live cells by DH was reported in the last decade (27). Recently, the tomographic phase-contrast microscopy (TPM) was applied for LD 3D imaging within mammalian cells (28, 29) and microalgae (30), for 4D tracking of the LDs dynamics in live hepatocytes (31), and for recording time-lapses of living foam cells (32). Nevertheless, the up-to-date available label-free techniques allow to investigate LDs only in static, adherent cells, strongly limiting both throughput and reliability of the information regarding the LD spatial organization, the latter being significantly affected by the cell culture mode.…”

Section: Introductionmentioning

confidence: 99%

Lipid droplets 3D full measurement by holographic in-flow tomography

Pirone

Sirico

Miccio

et al. 2021

Preprint

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The most recent discoveries in the biochemical field are highlighting the increasingly important role of lipid droplets (LDs) in several regulatory mechanisms in living cells. LDs are dynamic organelles and therefore their complete characterization in terms of number, size, spatial positioning and relative distribution in the cell volume can shed light on the roles played by LDs. Until now, fluorescence microscopy and transmission electron microscopy are assessed as the gold standard methods for identifying LDs due to their high sensitivity and specificity. However, such methods generally only provide 2D assays and partial measurements. Furthermore, both can be destructive and with low productivity, thus limiting analysis of large cell numbers in a sample. Here we demonstrate for the first time the capability of 3D visualization and the full LD characterization in high-throughput with a tomographic phase-contrast flow-cytometer, by using ovarian cancer cells and monocyte cell lines as models. A strategy for retrieving significant parameters on spatial correlations and LD 3D positioning inside each cell volume is reported. The information gathered by this new method could allow more in depth understanding and lead to new discoveries on how LDs are correlated to cellular functions.

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Regulation of lipid droplets in live preadipocytes using optical diffraction tomography and Raman spectroscopy

Cited by 14 publications

References 27 publications

Single‐shot quantitative phase imaging as an extension of differential interference contrast microscopy

Single‐shot quantitative phase imaging as an extension of differential interference contrast microscopy

Effect of Lauric vs. Oleic Acid-Enriched Diets on Leptin Autoparacrine Signalling in Male Mice

Lipid droplets 3D full measurement by holographic in-flow tomography

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