Preliminary exploration of the Δ conductivity quantification method for cell membrane permeability

doi:10.3969/j.issn.1004-1524.20240719

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (6): 1344-1352.DOI: 10.3969/j.issn.1004-1524.20240719

• Biosystems Engineering • Previous Articles Next Articles

Preliminary exploration of the Δ conductivity quantification method for cell membrane permeability

WANG Zhong(), YANG Hongbing, YANG Fan, CHEN Yifan, HOU Xiaomin^*()

College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, Shandong, China

Received:2024-08-07 Online:2025-06-25 Published:2025-07-08

Abstract

Abstract:

To enhance the reproducibility and comparability of cell membrane permeability assays and further quantify cell membrane permeability based on these improvements, a novel method for quantifying cell membrane permeability was successfully developed through investigating wheat leaf cell membrane permeability: the Δ conductivity quantification method for cell membrane permeability (abbreviated as the Δ conductivity method). The Δ conductivity method defines the unit of cell membrane permeability as the conductivity value of the extracellular solution leaked from 1 g of biomass with intact cell membranes after immersion in 1 L of pure water for 1 h under specific pressure. The calculation formula is: P=[(σ_e-σ_i)×V]/(m×R×t), where, P represents cell membrane permeability (with the unit being μS·cm^-1·L^-1·g^-1·h^-1), σ_e and σ_i represent the electrical conductivity at the end time point and the initial time point, V represents solution volume, m represents dry weight of sample, R represents intact rate of cell membrane, t represents consumption time. By comparing with the two existing common methods(air extraction method and soaking method), the results demonstrated that the three methods showed consistent conclusions and comparable reliability in qualitative research. However, the Δ conductivity method exhibited significantly superior repeatability compared with the air extraction method and soaking methods, proving that the Δ conductivity method can accurately and reliably quantify cell membrane permeability. The Δ conductivity method establishes a foundation for quantifying cell membrane permeability, making it potentially feasible for cell membrane permeability to serve as a hallmark biological indicator.

Key words: cell membrane permeability, Δ, conductivity method, quantify, reproducibility, reliability

CLC Number:

Q945
S512.1

WANG Zhong, YANG Hongbing, YANG Fan, CHEN Yifan, HOU Xiaomin. Preliminary exploration of the Δ conductivity quantification method for cell membrane permeability[J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1344-1352.

Figures/Tables 7

Fig.1 Schematic diagram of wheat leaf sampling A, Wheat seedlings at the two-leaf-with-one-heart stage; B, Mid-section of leaf.

Fig.2 Example diagram for selecting points σi and σe A, Prior to selecting points; B, After selecting points.

Fig.3 Self-made conductivity detection system A, Physical image of a self-made conductivity detection system; B, Structural diagram of a self-made conductivity detection system. 1, Vacuum pump; 2, KF adapter for air tubing connection; 3, Distribution box; 4, Power supply; 5, Check valve; 6, Vent valve; 7, Digital electrical contact pressure gauge (for vacuum pump control); 8, 100 L Vacuum tank; 9, Air tube connector; 10, Multi-path air tube splitter; 11, Hanging hole; 12, Stainless steel clamp (for securing vacuum tubes to wooden boards); 13, Nylon cable tie (for securing vacuum tubes to wooden boards); 14, Modified sample tube (5 mL centrifuge tube with multiple Φ 1 mm holes drilled); 15, Φ 1 mm Zinc-coated steel wire (wound and secured around the sample tube); 16, Modified 5 mL centrifuge tube cap (sheared, drilled, and adapted); 17, Ball valve with dual gas tube connections; 18, Thickened O-ring rubber seal; 19, Vacuum tube valve; 20, Leici DDSJ-318 conductivity meter; 21, Leici DJS-1D conductivity electrode; 22, Leici T-818-6 temperature electrode; 23, Computer; 24, USB data cable (for connecting conductivity meter to computer); 25, Silicone tube (Φ 6 mm×12 mm); 26, Cable for digital electrical contact pressure gauge; 27, Vacuum tube (modified for chromatography column); 28, Magnet.

Fig.4 Example of the statistical process for intact rate of cell membrane A, Scan image; B, ImageJ-generated total area graph; C,ImageJ-generated stained area graph.

Table 1 The statistical table of single-factor ANOVA test for cell membrane permeability measured by three methods

方法 Method	分组名称 Group name	CMP	CMP_mean	P
抽气法	0.6gF	18.007 9±1.898 9	17.381 9±1.283 0	0.466 5
Air extraction method	0.5gF	17.514 6±0.993 4
	0.4gF	16.623 1±0.716 7
	0.6gS	13.146 1±0.323 0	13.546 1±0.925 8	0.001 0
	0.5gS	14.699 5±0.151 2
	0.4gS	12.792 8±0.463 1
浸泡法	0.6gF	15.078 2±0.999 3	16.701 5±1.659 0	0.001 5
Soaking method	0.5gF	18.640 5±0.418 3
	0.4gF	16.385 7±0.306 4
	0.6gS	10.522 8±0.233 9	13.936 0±2.788 0	0.000 1
	0.5gS	16.740 1±0.334 9
	0.4gS	14.545 3±1.048 4
Δ电导率法	0.6gF	2.610 1±0.249 1	2.615 7±0.149 8	0.566 8
Δ conductivity method	0.5gF	2.690 2±0.064 0
	0.4gF	2.546 8±0.090 5
	0.6gS	2.105 9±0.100 7	2.086 0±0.163 7	0.873 8
	0.5gS	2.111 7±0.300 3
	0.4gS	2.040 3±0.046 3

Table 2 Statistical table of one-way ANOVA analysis for biological replication levels using three different methods

方法 Method	个案数 Number of cases	F/S平均值 Average of F/S ratio	P
抽气法Air extraction method	27	1.287 5±0.114 0	0.235 3
浸泡法 Soaking method	27	1.225 8±0.162 8
Δ电导率法 Δ conductivity method	27	1.260 4±0.113 9

Table 3 Statistical table of one-way ANOVA analysis for technique replication level using three methods

方法 Method	分组名称 Group name	个案数 Number of cases	F/S平均值 Average of F/S ratio	P
抽气法Air extraction method	0.6g	9	1.370 4±0.128 4	0.001 1
	0.5g	9	1.191 6±0.059 5
	0.4g	9	1.300 5±0.062 9
浸泡法Soaking method	0.6g	9	1.433 4±0.086 7	<0.000 1
	0.5g	9	1.113 8±0.029 1
	0.4g	9	1.130 3±0.070 2
Δ电导率法	0.6g	9	1.241 3±0.115 1	0.625 3
Δ conductivity method	0.5g	9	1.291 0±0.158 8
	0.4g	9	1.248 7±0.045 5

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Preliminary exploration of the Δ conductivity quantification method for cell membrane permeability

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