Simple sets for digital microphotography used and tested in the study of microorganisms

Four simple sets for digital microphotography are described that have been tested with the Carl Zeiss Jena, Meopta Prague, Lambda Prague, and LOMO Sankt Petersburg microscopes and with DSLR Nikon D 70 and Nikon D 300 cameras. They permit precise image focusing in the camera using a prism Zeiss. The sets make use of commonly available extensions Zeiss, Praktica and reductions Nikon??Praktica manufactured by ROWI (without a lens) or HAMA (with a lens). An extension has further been designed and manufactured for connecting the DSLR Nikon D 300 camera fitted with the HAMA reduction (only with a lens) and a focusing extensible prism with Zeiss Jena light measurement. It permits a precise image focusing of low light intensity objects (autofluorescence or low-contrast or moving objects when using positive or negative phase contrast). The sets are applicable to all microscopes constructed according to German DIN industrial standards.     

Greatness and tribulations of Zeiss and Leitz, two famous German optic companies III. Zeiss Ikon and elimination of Emanuel Goldberg

Gathering archival documents to trace the history of the Zeiss company presents no difficulty : they are abundant… except for a period from 1932 to 1945, systematically ignored, and that corresponds to the Nazi period. On the website Zeiss Historica, among the outstanding personalities of the Zeiss company, we note that, for Professor Emanuel Goldberg, the web page ? is still under development but an early picture of the professor is available. ?. But fortunately, Mickael Buckland, a Professor at the UC Berkeley School of Information brought the life and the work of Emanuel Goldberg to light. Thanks to him, his works and innovations, who had disappeared from our cultural and scientific heritage, return to light after being erased during fifty years. Goldberg had published dozens of articles, obtained patents, developed cameras, microdots, movie cameras, and he designed what he called a "Statistical Machine ", the first electronic document retrieval machine. In France, if this rediscovery was made known to the world of information science, it has not had the impact it deserved in the scientific world. Therefore it is time to reconstruct his career and his work, and to analyse the reasons why some attempted to erase definitively his name and memory.     

Turks in the New Germany

The Turkish community in Germany is fractured along ethnic, class, religious, and generational lines, although the practice of reciprocity provides stability and continuity in ethnic identification. Turks are also categorized by German discourses, which shifted after reunification, incorporating Turks into an anxiety-laden east-west problematic. Turkish responses to antiforeigner violence reflect ethnicity both as category and as practice: withdrawal behind communal boundaries or creation of a transnational creole ethnic self around the practice of reciprocity.     

Multiple excitation confocal analysis of targets in nuclei of cytogenetic preparations

OBJECTIVE: To demonstrate that cellular preparations requiring color analysis of different domains stained by molecular cytogenetic methods (fluorescence in situ hybridization) can be processed by spectral analysis of fluorescent emissions by either factor analysis of medical image sequences (FAMIS) or a META confocal configuration to isolate fluorescent probes. STUDY DESIGN: Three-dimensional sequences of images obtained by spectral analysis in a META confocal microscope (Carl Zeiss SAS, Jena, Germany) were analyzed by META processing and the FAMIS algorithm, which provides factor curves. META and factor images were then the result of image-processing methods that cover emission spectra. RESULTS: Factor curves and factor or META images can help to analyze targets inside nuclei. CONCLUSION: It is possible to process preparations containing numerous spots on different colors to differentiate stained targets and to improve visualization and detection.     

Repeatability Using Automatic Tracing with Canon OCT- HS100 and Zeiss Cirrus HD-OCT 5000

Background

Optical coherence tomography (OCT), can be used in clinical practice to provide high resolution cross-sectional images of the retina, optic disc and macula structure. These measurements can be useful for early detection, diagnosis, monitoring and treatment guidance for retinal diseases. Therefore, repeatability of measurements in OCT is of great importance.

Methods

Macula and optic disc parameters from the right eye of 30 healthy subjects were obtained twice with the Canon OCT-HS100 and Zeiss Cirrus HD-OCT 5000. Repeatability was evaluated by use of the coefficient of repeatability (CR) and the coefficient of repeatability as a percentage of the mean (CR%), and the obtained measurements were compared between the instruments.

Results

CR% of optic disc parameters ranged between 0.90 and 22.22% and 0.00 and 16.00% with the Canon and Zeiss OCT respectively. For macular parameters CR% ranged between 0.62 and 2.81% and 0.99 and 1.81% with the Canon and Zeiss OCT respectively. No statistical difference could be found when comparing the CR of all macular and disc measurements between the instruments. Compared to our previously published data repeatability has significantly improved with the inclusion of automatic tracking systems with both the Canon and Zeiss OCT.

Conclusion

Automatic tracking function improves repeatability in both Canon OCT-HS100 and Zeiss Cirrus HD-OCT 5000. However, measurements generated by the two instruments are still not interchangeable.     

Is agNOR and DNA ploidy analysis useful for evaluating thyroid neoplasms?

OBJECTIVE: To correlate the subjective AgNOR counting method and DNA content with histologic diagnoses of thyroid cancer and invasion. STUDY DESIGN: Eighty-one consecutive cases of thyroid carcinoma were selected for DNA and AgNOR analysis. The diagnoses were: papillary carcinoma (n = 40), follicular carcinoma (n = 31), Hürthle cell adenocarcinoma (n = 4), and undifferentiated carcinoma (n = 6). Seven normal thyroids were used as controls. DNA quantitative measurement was performed with Vidas 2.0 software (Kontron Bildanalyse, Munich, Germany) connected to an MPM 210 photometer microscope (Carl Zeiss, Oberkochen, Germany). The DNA index was obtained using histograms. Counting the NORs was performed by subjectively counting the NORs in 200 malignant cells. RESULTS: DNA ploidy analysis showed all Hürthle cell adenocarcinomas, 21 (67%)follicular tumors, 23 (57%) papillary tumors and 4 (67%) undifferentiated carcinomas to be aneuploid. DNA analysis correlated with histologic type of the tumor (p = 0.032). There was no statistical significance to the AgNOR counting variables studied. Statistical analysis showed correlation between ploidy and histologic diagnosis, but not AgNOR counting, to have prognostic value. CONCLUSION: DNA ploidy is more useful than subjective counting of NORs as an adjunct method for thyroid lesion analysis.     

Prostaglandin E2 receptor subtypes, EP1, EP2, EP3 and EP4 in human and mouse ocular tissues--a comparative immunohistochemical study

The purpose of the present study was to compare the localization of prostaglandin E(2) receptor subtypes in normal human and mouse ocular tissues. Paraffin embedded sections of normal human and mouse (129 Sv/Ev) eyes were treated with EP(1), EP(2), EP(3) and EP(4) specific antibodies and subsequently incubated with Alexa Fluor secondary antibody (Ex/Em=555/571) to detect the presence of EP receptor proteins. Fluorescence of the localized antibodies was visualized in a Carl Zeiss Microscope (Axiovert 200) and photographed using Carl Zeiss Axiocam camera. In mice EP(1) and EP(3) receptor subtypes were only moderately expressed, EP(3) receptor expression being almost negligible. In human cornea and iris ciliary body, EP(1) and EP(3) receptors were prominently expressed. EP(4) receptor was expressed moderately in human and mouse ocular tissues. EP(2) receptor was the most prominently and abundantly expressed receptor in both human and mouse ocular tissues. It is concluded that the pattern of the distribution of EP receptor subtypes in the ocular tissues are similar in human and mouse. Thus, 129 Sv/Ev strains of mice would make an appropriate animal model for studying the ocular pathophysiological roles of prostaglandin receptor agonists.     

The work by Giulio Ceradini in explaining the mechanism of semilunar cardiac valve function

Using an excised pig heart preparation with tubes, a manometer, and a visualizing apparatus, Giulio Ceradini, an Italian physiologist working in the years of 1871-1872 in Carl Ludwig's famous laboratory in Leipzig, Germany, illustrated the mechanism of closure of the semilunar valves. He was the first to conceive that the closure of the heart valves depends not on a static back pressure nor upon eddies but is primarily the consequence of the decelerated systolic efflux. This pioneer research of Ceradini was first published in German in 1872 (4). The purpose of the present report is to revisit Ceradini's pioneering experiments and his interpretation of heart valve closure, which remains as true as it was in 1872.     

Rapid continuous monitoring of enzyme activity in tissue sections: experience with the M85A and Zeiss UMSP-30 systems

We have previously described methods for the continuous monitoring of formazan deposition in tissue sections with a system based on the Vickers M85A microdensitometer. However, this instrument only allows the monitoring of a single field in each section. We have now developed a new system based on the Zeiss UMSP-30 microspectrophotometer. This machine is entirely computer controlled and by virtue of its fast-scanning stage allows the rapid (less than 0.5 s) sequential monitoring of multiple fields (up to 35) in each section. Thus a number of cell types may be studied simultaneously and work which used to take a full working day with the M85A system now can be performed in 45 min. As with the M85A the Zeiss system has full capability for data analysis (i.e. calculation of initial velocity rates, etc.). We have found that continuous monitoring of tissue sections by microdensitometry is a precise, sensitive and biochemically valid method of studying enzyme activity within the cellular matrix.     

Morphometric analysis of AgNORs in thin-layer, liquid-based liver specimens

OBJECTIVE: To detect argyrophilic nucleolar organizer regions (AgNORs) in ThinPrep (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) liver fine needle aspiration (FNA) specimens and to define the diagnostic value of their quantitative analysis in the evaluation of hepatic lesions. STUDY DESIGN: ThinPrep liquid-based FNA biopsy specimens from 49 malignant and benign liver lesions were resampled, fixed in 95% ethanol and stained with the AgNOR technique in accordance with the 1-step colloid method. The specimens included 11 benign and 38 malignant lesions (23 poorly differentiated hepatocellular carcinomas [HCCs] and 15 poorly differentiated metastatic adenocarcinomas [MCs]). Morphometric analysis was performed using a Zeiss Axiolab microscope (Carl Zeiss GmbH, Jena, Germany) with a mechanical stage fitted with a Sony-iris CCD videocamera (Tokyo, Japan). The videocamera was connected to a Pentium III P/C (Intel Corp., Santa Clara, California, U.S.A.) loaded with the appropriate image analysis software. The measurements were performed with ImageScan software (Jandel Scientific, Erkrath, Germany). The number of AgNORs per nucleus (NN) and the total area per nucleus occupied by AgNORs (AR) were calculated semiautomatically. Statistical analysis was performed using the SPSS software package (Chicago, Illinois, U.S.A.). RESULTS: The least significant deviance test for multiple comparisons revealed that NN differed significantly between the 3 groups of samples examined (P < .0001). The mean NN values in HCCs and MCs were significantly different (P < .0001). Logistic regression model demonstrated that as NN increased, the probability of a MC diagnosis decreased (<4%). AR values were different at a statistically significant level only between benign and malignant specimens (P = .00006), not between HCCs and MCs (P = .933). CONCLUSION: Quantitative analysis of AgNORs in ThinPrep specimens could be a diagnostically useful method in liver disease.     

From Ethical Exceptionalism to Ethical Exceptions: The Rule and exception Model and the Changing Meaning of Ethics In German Bioregulation

Germany is an interesting case with respect to the governance of reprogenetics. It has a strong profile in the technosciences and high aims regarding the global bioeconomy, yet her regulation of human genetics, reproductive medicine and embryo research has for a long time been rather restrictive. German biopolitical exceptionalism has often been explained by reference to Catholicism and the legacy of the Nazi past. The Germans, so goes the common story, have learnt the lessons of history and translated them into unconditional respect for human dignity, which in turn translates into unconditional protection of human life, including the human embryo, and the firm repudiation of any eugenic distinction between ‘life worth to live’ and ‘life not worth to live’. This, however, is not the whole story. Alongside deontological strictness we find another strand of governing body politics and reprogenetics in Germany, the rule‐and‐exception model, running from the mid‐1970s abortion law via the 2002 Stem Cell Act to the 2011 regulation of pre‐implantation genetic diagnosis. In contrast to the former, that strongly draws on Kant and his concept of human dignity, the latter bears resemblances to Carl Schmitt's concept of state of exception. The article will show that the rule‐and‐exception model builds the exception into the rule and transforms the meaning and mandate of ethics, namely from safeguarding ethical standards to deciding about the exception. Given that the exception has now tended to become the rule, the question is whether the lessons of history will govern German reprogenetics for much longer.     

An American in Paris and the origins of the stereomicroscope

A century ago, Horatio S. Greenough, an American living in Paris, persuaded the firm of Carl Zeiss to construct the first low-power stereomicroscope. Fitted by Zeiss with Porro prisms for image erection, this instrument became the ancestor of all stereoscopic dissection microscopes (binoculars) now in use. On the basis of original documents and earlier publications, the present Centennial Essay traces both the history of the Greenough stereomicroscope and the scantily documented life of its inventor, including his probable descent from a family of famous New England sculptors. The aim is to provide some historical background for a truly timeless instrument that was fundamental to developmental biology, beginning almost from the days of its proclamation by Wilhelm Roux.     

An improved method for the automated enumeration of fluorescently labelled bacteria in human faeces

The study aimed to improve microscopy-based automated recognition of faecal bacterial cells labelled with 16S rRNA-targeted oligonucleotides and 4',6-diamidino-2-phenylindole (DAPI). Based on the software KS400 (Carl Zeiss Vision, Hallbergmoos, Germany), designed for automising microscopy-based image capture and image analysis, a routine was developed that affords the recognition of doubly stained bacteria and the rejection of artefacts. The accuracy of the automated enumeration was investigated by comparing the resulting data with those obtained by manual counting. The newly developed method was subsequently used to compare the total bacterial counts in human faecal samples using the domain specific probe Eub338 alone and a mixture of 5 domain-specific probes, respectively. Faecal samples from 90 healthy volunteers were analysed. The cell counts obtained with Eub338 were 10% lower than those obtained with the probe mixture. Since the cells detected with the probe mixture covered a wide range of signal intensities, a dynamic analysis routine was developed to effectively detect the whole range of bright to weak signals within the same image, while at the same time reliably rejecting artefacts.     

Use and misuse of nitrogen in agriculture: the German story

Nitrogen (N) fertilization in agriculture has been discussed controversially in Germany for almost two centuries. The agronomist Carl Sprengel, who published his theory on the mineral nutrition of plants in 1828, advocated the use of mineral N fertilizers. Chemist Justus von Liebig, on the other hand, vehemently denied around 1850 the need for N fertilization. Although it soon became evident that Sprengel was right and Liebig was wrong, not much synthetic N fertilizer was used in German agriculture until around 1915, when the Haber-Bosch technique enabled the commercial production of NH3. The use of N fertilizers since then has grown, especially since 1950. To increase agricultural productivity, German governments have promoted, directly and indirectly, the use of N in crop and in animal production. Unfortunately, it was overlooked that N surpluses in agriculture increased rapidly; around 1980 they amounted yearly to more than 100 kg ha(-1). The extensive use of N in agriculture is causing environmental damage and is contributing substantially to the external costs of present agriculture. The main N compounds that affect the environment are N2O, NH3, and NO3. These compounds are considered to contribute one third to the external costs of agriculture. Additionally, the high rate of human intake of animal proteins and lipids has adversely affected the health of the country's population. Fundamental corrections in German farm policy appear inevitable.     

Secular changes are different in distinct subgroups of the growing population

The change of living conditions in East Germany after the German reunification in 1990 led to intensive secular changes in growth and development. Anthropometric data of height, weight, BMI and thickness of the subcutaneous fat layer are compared from two cross-sectional samples of German children and adolescents aged 6 to 18 years. The first sample was measured around 1989 and the second 10 years later around 1999. Both samples contain children and adolescents from the urban as well as from the rural population. The secular changes reported here are based on the comparison of medians of height, weight, BMI and subcutaneous fat layer thickness calculated for yearly age categories. Subsequently, medians of the 10% border categories are compared. These border categories contain the 10% smallest, lightest, slimmest or leanest subjects and the 10% tallest, heaviest, most corpulent or most obese subjects respectively of a yearly age category. The young generation shifted between 1989 and 1999 to a taller stature in the small as well as in the tall category. The secular pattern of measurements, which mark corpulence, is sex-specific and differs from that of length measurements. In general the light or lean subjects changed only little, however, there are clear age- and sex-specific changes in the upper border categories. Weight and BMI increased markedly in the upper border categories in young preadolescent children, but did not change much in adolescent boys of these category aged 15 and 16 and decreased in adolescent girls. Also the subcutaneous fat layer increased in the upper border category in preadolescent children, but decreased in adolescent boys and girls.     

Precision and Agreement of Corneal Power Measurements Obtained Using a New Corneal Topographer OphthaTOP

Purpose

To evaluate repeatability and reproducibility of anterior corneal power measurements obtained with a new corneal topographer OphthaTOP (Hummel AG, Germany) and agreement with measurements by a rotating Scheimpflug camera (Pentacam HR, Oculus, Germany) and an automated keratometer (IOLMaster, Carl Zeiss Meditec, Germany).

Methods

The right eyes of 79 healthy subjects were prospectively measured three times with all three devices. Another examiner performed three additional scans with the OphthaTOP in the same session. Within one week, the first examiner repeated the measurements using the OphthaTOP. The flat simulated keratometry (Kf), steep K (Ks), mean K (Km), J₀, and J₄₅ were noted. Repeatability and reproducibility of measurements were assessed by within-subject standard deviation (Sw), repeatability (2.77 Sw), coefficient of variation (CoV), and intraclass correlation coefficient (ICC). Agreement between devices was assessed using 95% limits of agreement (LoA).

Results

Intraobserver repeatability and interobserver and intersession reproducibility of all measured parameters showed a 2.77 Sw of 0.29 diopter or less, a CoV of less than 0.24%, and an ICC of more than 0.906. Statistically significant differences (P<0.001) were found between the parameters analyzed by the three devices, except J₀ and J₄₅. The mean differences between OphthaTOP and the other two devices were small, and the 95% LoA was narrow for all results.

Conclusions

The OphthaTOP showed excellent intraobserver repeatability and interobserver and intersession reproducibility of corneal power measurements. Good agreements with the other two devices in these parameters were found in healthy eyes.     

Three-dimensional imaging of tumor angiogenesis

OBJECTIVE: To three-dimensionally visualize the microvessel environment of tumor angiogenesis by confocal laser scanning microscopy (CLSM). STUDY DESIGN: To reveal underlying mechanisms of tumor angiogenesis, a 7, 12-dimethylbenz(a) anthracene-induced rat cancer model was used. For demonstrating tumor vasculature, fluorescence injection method (FITC-conjugated gelatin solution) was employed. FITC gelatin was injected into the left ventricle of the rat heart. After complete perfusion, the mammary glands were resected, fixed under ice cold conditions and subjected to immunohistochemistry (IHC) for tumor cells. The LSM-410 (Carl Zeiss, Jena, Germany) was employed on thick sections (300-2,000 microns) to elucidate detailed microvessel networks (MVN) and tumor cells. RESULTS: Tumor vasculature on thick sections was clearly detected by CLSM at the maximum focus depth of 2,000 microns. Three-dimensional (3-D), reconstructed images of normal mammary glands showed regular and linear MVN. In DMBA-induced mammary cancer, vascular density of MVN was markedly increased and showed an anastomosing, irregular MVN pattern. Furthermore, focal segmentation and tortuous, branching patterns of microvessels were also seen. CONCLUSION: Application of the fluorescence injection method and IHC using CLSM was very useful for studying the 3-D relationship between tumor angiogenesis and neoplastic epithelial changes. These results suggest that application of this technique is ideal for studying 3-D imaging of tumor angiogenesis.     

On Carl Gegenbaur’s theory on head metamerism and the selection of taxa for comparisons

Summary After the rejection of the idealistic “vertebral theory of the skull,” the basic idea of the theory had been taken up again by the German morphologist Carl Gegenbaur. He proposed a metameric ordering system of the vertebrate head based on a comparative investigation of selachians. This “segmentation theory of the vertebrate head” had a stimulating effect on comparative anatomy. Traces of the basic ideas are still inherent in the biological sciences. Despite the fact that Carl Gegenbaur proposed a rather phylogenetic approach, his systematic investigations remained on a typological level. It is argued herein, that the reason for that was his inability to perform his comparisons independent of a rather intuitive a priori hypothesis about phylogenetic relationships, thus leaving no possibilities for testing homologies. This is exemplified by Carl Gegenbaur’s choice of species for comparative work as well as his use of these species in comparative work. Choosing species for comparisons is discussed.     

No More Foreigners? The Remaking of German Naturalization and Citizenship Law, 1990–2000

Since the beginning of the 1990s, Germany has been taking controversial policy measures that had been quite unimaginable, even in the 1980s. On the one hand, regulations for asylum seekers and ethnic Germans have gradually become restrictive in nature. On the other hand, changes to the Citizenship and Nationality Law of 1913 (Reichs und Staatsangehörigkeitsgesetz) and the Alien Act of 1965 (Ausländergesetz) have dramatically altered the naturalization process. Before the passage of the new German citizenship law in 1999, the acquisition of citizenship in Germany was solely based on the principle of descent (jus sanguinis). This paper attempts to explain why German government altered its citizenship policy and introduced birthright citizenship. It reviews the post-war contextual factors that have shaped the debate on citizenship and asks whether it is external or internal factors, that is international-level norms and institutions or domestic politics, that led to the change. If domestic politics can fully explain the adoption of the new citizenship law, what domestic forces played an important role? Was state action taken in the name of protecting “the national interest” or was it party politics? What role did societal forces play in the making of new citizenship policy? Did the act seek to mitigate anti-foreigner sentiment growing in Germany since the 1990s? Using archival and documentary research and secondary data on socio-economic and demographic trends in Germany, this paper concludes that domestic politics explain the changes in the citizenship law. I argue that granting and withholding national citizenship, including birthright citizenship, is exclusively in the hands of a nation state. Considering a purely ethnocultural understanding of nation-hood now includes legal and territorial components, the new German citizenship law is a major reform at the both theoretical and conceptual level. However, the practical consequences of the new naturalization process remain to be seen. The main question for the future research agenda is whether the new naturalization and citizenship laws will bring full integration of foreigners living in German society.     

Effect of chitosan on the electrophoretic motility of thymocytes

An increase of contact interaction between macrophages and thymocytes in the presence of polysaccharide chitosan was suggested to be due to the change of surface charge of interacting cells. It was found that incubation of thymocytes in the presence of chitosan leads to the change of their charge in the positive direction. The measurement of electrophoretic mobility of the cells was carried out on " Parmoquant 2" (Carl Zeiss Jena, DDR). The change of electrophoretic mobility increases with decrease of pH, increase of chitosan concentration and decrease of the cell concentration in the medium. This phenomenon is probably due to absorption of positively charged molecules of chitosan on negatively charged cell surface.