Abstract
The development of the segment-flow technology has considerably extended the application range of miniaturized calorimeters. The transport of samples in aqueous µL segments by a water-immiscible carrier liquid enables the investigation of aggregated, complex biological samples in flow-through. The protection from contaminations of the fluidics of the calorimeter by the carrier liquid is an excellent precondition for the design of efficient automatic experiment protocols. In this paper, a new operation mode of this technique is presented, which allows the measurement of fast transient heat-dissipating processes using a chip calorimeter. Heat dissipation is initiated by the merging of separate µL-sample-segments arriving successively at the measuring chamber. The potential of the segment fusion technique was demonstrated here using two different applications. First, we showed the use of this method for an automatic screening of the strength of receptor–substrate interactions by the measurement of the heat of the adduct formation of carbohydrate octyl ß-d-glucopyranoside and three different receptor molecules. Next, we used the method for the determination of transient heat effects in the seconds range allowing the investigation of metabolic processes in Trypanosoma cruzi cells induced by Ca2+ ions.
Similar content being viewed by others
Availability of data and materials
At the authors.
References
Lerchner J, Wolf G, Auguet C, Torra V. Accuracy in integrated circuit (IC) calorimeters. Thermochim Acta. 2002;382:65–76.
Lerchner J, Wolf A, Wolf G, Fernandez I. Chip calorimeters for the investigation of liquid phase reactions: design rules. Thermochim Acta. 2006;446:168–75.
Wang S, Sha X, Yu S, Zhao Y. Nanocalorimeters for biomolecular analysis and cell metabolism monitoring. Biomicrofluidics. 2020;14:011503.
Hartmann T, Barros N, Wolf A, Siewert C, Volpe PL, Schemberg J, Grodrian A, Kessler E, Hänschke F, Mertens F, Lerchner J. Thermopile chip based calorimeter for the study of aggregated biological samples in segmented flow. Sens Actuators B. 2014;210:460–8.
Lerchner J, Volpe POL, Lanaro C, Fertrin KY, Costa FF, Albuquerque DM, Haenschke F, Mertens F. A chip calorimetry based method for the real-time investigation of red blood cell sickling. J Therm Anal Calorim. 2019;136:771–81.
Lerchner J, David KA, Unger FT, Lemke K, Förster T, Mertens F. Continuous monitoring of drug effects on complex biological samples by segmented flow chip calorimetry. J Therm Anal Calorim. 2017;126(3):1307–17.
Wolf A, Hartmann T, Bertolini M, Schemberg J, Grodrian A, Lemke K, Foerster T, Kessler E, Haenschke F, Mertens F, Paus R, Lerchner J. Toward high-throughput chip calorimetry by use of segmented-flow technology. Thermochim Acta. 2015;603:172–83.
Wang B, Jia Y, Lin Q. A microfabrication-based approach to quantitative isothermal titration calorimetry. Biosens Bioelectron. 2016;78:438–46.
Lee W, Fon W, Axelrod BW, Roukes ML. High-sensitivity microfluidic calorimeters for biological and chemical applications. PNAS. 2009;106:15225–30.
Docampo R, Vercesi AE. Ca2+ transport by coupled Trypanosoma cruzi mitochondria in situ. J Biol Chem. 1989;264:108–11.
Docampo R, Vercesi AE. Characteristics of Ca2+ transport by Trypanosoma cruzi mitochondria in situ. Arch Biochem Biophys. 1989;272:122–9.
Lerchner J, Maskow T, Wolf G. Chip calorimetry and its use for biochemical and cell biological investigations. Chem Eng Proc. 2008;47:991–9.
Köhler JM, Henkel T, Grodrian A, Kirner T, Roth M, Martin K, Metze J. Digital reaction technology by micro segmented flow—components, concepts and applications. Chem Eng J. 2004;101:201–16.
Budden M, Schneider S, Groß GA, Kielpinski M, Henkel T, Köhler JM. Splitting and switching of microfluid segments in closed channels for chemical operations in the segment-on-demand technology. Chem Eng J. 2013;227:166–73.
O’Neill MAA, Beezer AE, Vine GJ, Kemp RB, Olomolaiye D, Volpe POL, Oliveira D. Practical and theoretical consideration of flow-through microcalorimetry: determination of “thermal volume” and its flow rate dependence. Thermochim Acta. 2004;413:193–9.
Stapf M, Seichter W, Mazik M. Cycloalkyl groups as subunits of artificial carbohydrate receptors: effect of ring size of the cycloalkyl unit on the receptor efficiency. Eur J Org Chem. 2020;31:4900–15.
Stapf M, Seichter W, Mazik M. Influence of intramolecular interactions on the binding properties of acyclic carbohydrate-binding agents. Unpublished results.
Lippe J, Seichter W, Mazik M. Improved binding affinity and interesting selectivities of aminopyrimidine-bearing carbohydrate receptors in comparison with their aminopyridine analogues. Org Biomol Chem. 2015;13:11622–32.
Lander N, Chiurillo MA, Docampo R. CRISPR/Cas9 technology applied to the study of proteins involved in calcium signaling in Trypanosoma cruzi. Methods Mol Biol. 2020;2116:177–97.
Chiurillo MA, Lander N, Bertolini MS, Storey M, Vercesi AE, Docampo R. Different roles of mitochondrial calcium uniporter complex subunits in growth and infectivity of Trypanosoma cruzi. MBio. 2017;8(3):e00574-17.
https://somapp.ucdmc.ucdavis.edu/pharmacology/bers/maxchelator/CaEGTA-NIST.htm.
Raal JD, Webley PA. Microflow calorimeter design. AIChE J. 1987;33:604–18.
Hill JO, Öjel G, Wadsö I. Thermochemical results for “tris” as a test substance in solution calorimetry. J Chem Thermodyn. 1969;1:111–6.
Mazik M. Recent developments in the molecular recognition of carbohydrates by artificial receptors. RSC Adv. 2012;2:2630–42.
Mazik M. Molecular recognition of carbohydrates by acyclic receptors employing noncovalent interactions. Chem Soc Rev. 2009;38:935–56.
Weber PC, Salemme FR. Applications of calorimetric methods to drug discovery and the study of protein interactions. Curr Opin Struct Biol. 2003;13:115–21.
Bianconi ML. Calorimetry of enzyme-catalyzed reactions. Biophys Chem. 2007;126:59–64.
Vercesi AE, Hoffmann ME, Bernardes CF, Docampo R. Regulation of intracellular calcium homeostasis in Trypanosoma cruzi: effects of calmidazolium and trifluoperazine. Cell Calcium. 1991;12:281–389.
Vercesi AE, Castilho RF, Kowaltowski AJ, Oliveira HCF, Souza-Pinto NC, Figueira TR, Busanello ENB. Mitochondrial calcium transport and the redox nature of the calcium induced membrane permeability transition. Free Rad Biol Med. 2018;129:1–24.
Lerchner J, Sartori MR, Volpe POL, Lander L, Mertens F, Vercesi AE. Direct determination of anaerobe contributions to the energy metabolism of Trypanosoma cruzi by chip calorimetry. Anal Bioanal Chem. 2019;411:3763–8.
Point R, Petit JL, Gravelle PC. Calorimetry for thermokinetic determinations. J Thermal Anal. 1979;17:383–95.
Acknowledgements
The authors are grateful to Raquel Negreiros for the help with preparation and manipulation of parasites.
Funding
This study was supported by the São Paulo Research Foundation (Fundação de Amparo à Pesquisa do Estado de São Paulo, FAPESP) visiting researcher Grant Nos. 2018/19976-1 to J.L/A.E.V., 2017/17728-8 to A.E.V., and postdoctoral fellowship to M.R.S./A.E.V (2017/05487-6).
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Johannes Lerchner and Marina Sartori. The first draft of the manuscript was written by Johannes Lerchner, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Lerchner, J., Sartori, M.R., Volpe, P.O. et al. Segment fusion chip calorimetry: a new method for the investigation of fast reactions. J Therm Anal Calorim 147, 2253–2263 (2022). https://doi.org/10.1007/s10973-021-10623-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10973-021-10623-7